Nanofilm and membrane compositions

ABSTRACT

Nanofilms useful for filtration are prepared from oriented amphiphilic molecules and oriented macrocyclic modules. The amphiphilic species may be oriented on an interface or surface. The nanofilm may be prepared by depositing or attaching an oriented layer to a substrate. A nanofilm may also be prepared by coupling the oriented macrocyclic modules to provide a membrane.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.10/359,894, filed Feb. 7, 2003 now U.S. Pat. No. 7,432,371, which claimsbenefit of U.S. Provisional Application No. 60/383,236, filed May 22,2002 and is a continuation-in-part of U.S. patent application Ser. No.10/226,400, filed Aug. 23, 2002, now abandoned, which is acontinuation-in-part of U.S. patent application Ser. No. 10/071,377,filed Feb. 7, 2002, now abandoned. Each of these applications isincorporated herein by reference for all purposes.

TECHNICAL FIELD

This invention relates variously to thin layer compositions of coupledoriented amphiphilic macrocyclic modules, nanofilms having particularpermeation properties, and nanofilms for filtration and separation. Thethin layer nanofilms may be applied to a variety of selective permeationprocesses.

BACKGROUND OF THE INVENTION

Nanotechnology involves the ability to engineer novel structures at theatomic and molecular level. One area of nanotechnology is to developchemical building blocks from which hierarchical molecules of predictedproperties can be assembled. An approach to making chemical buildingblocks or nanostructures begins at the atomic and molecular level bydesigning and synthesizing starting materials with highly tailoredproperties. Precise control at the atomic level is the foundation fordevelopment of rationally tailored synthesis-structure-propertyrelationships which can provide materials of unique structure andpredictable properties. This approach to nanotechnology is inspired bynature. For example, biological organization is based on a hierarchy ofstructural levels: atoms formed into biological molecules which arearranged into organelles, cells, and ultimately, into organisms. Thesebuilding block capabilities are unparalleled conventional materials andmethods such as polymerizations which produce statistical mixtures orconfinement of reactants to enhance certain reaction pathways. Forexample, from twenty common amino acids found in natural proteins, morethan 10⁵ stable and unique proteins are made.

One field that will benefit from nanotechnology is filtration usingmembranes. Conventional membranes used in a variety of separationprocesses can be made selectively permeable to various molecularspecies. The permeation properties of conventional membranes generallydepend on the pathways of transport of species through the membranestructure. While the diffusion pathway in conventional selectivelypermeable materials can be made tortuous in order to control permeation,porosity is not well defined or controlled by conventional methods. Theability to fabricate regular or unique pore structures of membranes is along-standing goal of separation technology.

Resistance to flow of species through a membrane may also be governed bythe flow path length. Resistance can be greatly reduced by using a verythin film as a membrane, at the cost of reduced mechanical strength ofthe membrane material. Conventional membranes may have a barrierthickness of at least one to two hundred nanometers, and often up tomillimeter thickness. In general, a thin film of membrane barriermaterial can be deposited on a porous substrate of greater thickness torestore material strength.

Membrane separation processes are used to separate components from afluid in which atomic or molecular components having sizes smaller thana certain “cut-off” size can be separated from components of largersize. Normally, species smaller than the cut-off size are passed by themembrane. The cut-off size may be an approximate empirical value whichreflects the phenomenon that the rate of transport of components smallerthan the cut-off size is merely faster than the rate of transport oflarger components. In conventional pressure-driven membrane separationprocesses, the primary factors affecting separation of components aresize, charge, and diffusivity of the components in the membranestructure. In dialysis, the driving force for separation is aconcentration gradient, while in electrodialysis electromotive force isapplied to ion selective membranes.

In all these methods what is required is a selectively permeablemembrane barrier to components of the fluid to be separated.

SUMMARY OF THE INVENTION

In one aspect, this invention relates to a nanofilm comprising coupledoriented amphiphilic macrocyclic modules. The modules of the nanofilmmay be coupled through reactive functional groups of the modules, or maybe coupled through a linker molecule. The coupling may be initiated bychemical, thermal, photochemical, electrochemical, or irradiativemethods.

In some variations, the thickness of a nanofilm is less than about 30nanometers, sometimes less than about 4 nanometers, and sometimes lessthan about 1 nanometer.

A nanofilm may have a filtration function that may be used to describethe species that pass through the nanofilm. A nanofilm may be permeableonly to a particular species in a particular fluid and species smallerthan the particular species. A nanofilm may have a molecular weightcut-off.

A particular nanofilm may have high permeability for certain species ina certain solvent. A nanofilm may have low permeability for certainspecies in a certain solvent. A nanofilm may have high permeability forcertain species and low permeability for other species in a certainsolvent.

A nanofilm barrier can be made up of layers of nanofilm. A spacing layermay be used between any two nanofilm layers. Spacing layers may includelayers of polymer, gel, and other substances.

A nanofilm may be deposited on a substrate, which in turn may be porousor non-porous. A nanofilm may have surface attachment groups, and may becovalently bonded to a substrate through surface attachment groups, orbonded to a substrate through ionic interactions.

In another variation, this invention relates to a method for filtrationcomprising using a nanofilm to separate components or species from afluid or solution.

In some instances, a nanofilm is composed of oriented macrocyclicmodules deposited on a substrate using a Langmuir trough. In othervariations, a nanofilm may be made up of coupled oriented amphiphilicmolecules and oriented amphiphilic macrocyclic modules.

In one variation, a nanofilm is composed of oriented amphiphilicmolecules coupled through a hydrophilic group.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A illustrates a Hexamer 1dh macrocvclic module, which may bereacted with diethyl malonimidate and at least one other macrocyclicmodule to yield the nanofilm illustrated in FIG. 1B.

FIG. 2A illustrates amphiphilic alkythiol molecules, which may bereacted to yield the nanofilm illustrated in FIG. 2B.

FIG. 3A illustrates an exemplary scheme for preparing a nanofilmcomprising amphiphilic methyl 2-amino(alkan)oate molecules, which may bereacted to yield the nanofilm illustrated in FIG. 3B.

FIG. 4A illustrates amphiphilic alkylamine molecules, which may bereacted with formaldehyde to yield the nanofilm illustrated in FIG. 4Bcomprising dialkylmethandiamine units. FIG. 4C illustrates amphiphilicalkylamine molecules, which may be reacted with formaldehyde to yieldthe nanofilm illustrated in FIG. 4D comprising alkylamine polymers. FIG.4E illustrates amphiphilic alkylamine molecules, which may be reactedwith formaldehyde to yield the nanofilm illustrated in FIG. 4Fcomprising 1,3,5-trialkyl-1.3,5-triazinane units.

FIG. 5A illustrates a nanofilm, which may be attached to the substrateof FIG. 5B, showing examples of surface attachment groups, to yield thecomplex illustrated in FIG. 5C.

FIGS. 6A and 6B illustrate examples of the ellipsometric images of thepreparation of a nanofilm of Hexamer 1 dh.

FIG. 7 illustrates an example of isobaric creep of a nanofilm of Hexamer1 dh.

FIGS. 8A and 8B illustrate examples of FTIR spectra of the preparationof a nanofilm of Hexamer 1 dh.

FIG. 9 illustrates an example of the ellipsometric image of thepreparation of a nanofilm of Hexamer 1 dh.

FIGS. 10A, 10B and 10C illustrate example of ellipsometric images of thepreparation of a nanofilm of methylheptadecanoate attached to asubstrate.

FIGS. 11A and 11B illustrate examples of ellipsometric images of thepreparation of a nanofilm of N-octadecylacrylamide attached to asubstrate.

FIG. 12 illustrates a representation of the structure of a nanofilm ofHexamer 1 dh.

FIG. 13 illustrates a representation of the structure of a nanofilm ofOctamer 5jh-aspartic.

FIG. 14 illustrates the Langmuir trough area versus time for a nanofilmprepared from Hexamer 1jh-AC.

FIG. 15 illustrates an example of the ellipsometric image of a nanofilmprepared from Hexamer 1jh-AC.

FIG. 16 illustrates an example of the FTIR spectra of the preparation ofa nanofilm of Hexamer 1jh-AC.

FIGS. 17A and 17B show representations of examples of the structure ofembodiments of a hexamer macrocyclic module.

FIG. 18A shows an example of the Langmuir isotherm of an embodiment of ahexamer macrocyclic module.

FIG. 18B shows an example of the isobaric creep of an embodiment of ahexamer macrocyclic module.

FIG. 19A shows an example of the Langmuir isotherm of an embodiment of ahexamer macrocyclic module.

FIG. 19B shows an example of the isobaric creep of an embodiment of ahexamer macrocyclic module.

DETAILED DESCRIPTION OF THE INVENTION Macrocyclic Modules and NanofilmCompositions

In one aspect, this invention relates to nanotechnology in thepreparation of porous structures and materials having pores that are ofatomic to molecular size. These materials may have unique structureswhich repeat at regular intervals to provide a lattice of pores havingsubstantially uniform dimensions. The unique structures may have avariety of shapes and sizes, thereby providing pores of various shapesand sizes. Because the unique structures may be formed in a monolayer ofmolecular thickness, the pores defined by the unique structures mayinclude a cavity, opening, or chamber-like structure of molecular size.In general, pores of atomic to molecular size defined by those uniquestructures may be used for selective permeation or molecular sievingfunctions. Some aspects of nanotechnology are given in NanostructuredMaterials, J. Ying, ed., Academic Press, San Diego, 2001.

This invention further includes the rational design of molecules thatmay be assembled as building blocks for further assembly into largerspecies. Standardized molecular subunits or modules may be used fromwhich hierarchical molecules of predicted properties can be assembled.Coupling reactions can be employed to combine or attach modules indirected syntheses.

Nanotechnology involves the assembly of molecular building blocks toform intermediate size hierarchical molecules having built-indirectionality. Ideally, a nanoassembly begins with a set of synthonswhich may be assembled to make a module. Synthons are individualmolecules which are the primary starting material. The module may have aset of “arms” destined for interconnection with other modules. Modulesare covalently-bonded combinations of synthons. Modules may be used asbuilding blocks for larger molecular species, including uniquelystructured species and compositions. A wide variety of real-worldapplications, such as membranes or porous materials, may be derived fromnano-chemical tools, compositions and processes.

Molecular modules may be prepared from cyclic organic synthons. Synthonsmay be coupled or bonded together to form modules. For example, amacrocyclic module may be prepared with the cyclic organic synthonsR,R-1,2-transdiaminocyclohexane and 4-substituted 2,6-diformyl phenol.These synthons may be coupled to form a hexameric macrocyclic modulehaving the following arrangement:

where each R group may be different.

This hexameric module and others are useful as building blocks toprovide structures having certain controlled and predictable properties.Macrocyclic modules and amphiphilic macrocyclic modules prepared fromcyclic organic synthons are described in U.S. patent application Ser.Nos. 10/071,377 and 10/226,400, and in the PCT Application entitled“Macrocyclic module compositions” filed Feb. 7, 2003, hereinincorporated by reference. Examples of synthons, macrocyclic modules,and amphiphilic macrocyclic modules and their syntheses are furtherdescribed hereinbelow.

Examples of modules useful as building blocks are shown in Table 1.

TABLE 1 Examples of macrocyclic modules MODULE STRUCTURE Hexamer 1a

Hexamer 1dh

Hexamer 3j- amine

Hexamer 1jh

Hexamar 1jh-AC

Hexamer 2j- amine/ester

Hexamer 1dh- acryl

Octamer 5jh- aspartic

Octamer 4jh- acryl

Macrocyclic modules can be oriented on a surface by providing functionalgroups on the modules which impart amphiphilic character to the modules.For example, when the module is deposited on a hydrophilic surface,hydrophobic substituent groups or hydrophobic tails attached to themodule may cause the module to reorient on the surface so that thehydrophobic substituents are oriented away from the surface, leaving amore hydrophilic facet of the module oriented toward the surface.Examples of hydrophobic groups include lower alkyl groups, alkyl groupshaving 7, 8, 9, 10, 11, 12, or more carbon atoms, including alkyl groupswith 14-30, or 30 or more carbon atoms, substituted alkyl groups, arylgroups, substituted aryl, saturated or unsaturated cyclic hydrocarbons,heteroaryl, heteroarylalkyl, heterocyclic, and corresponding substitutedgroups. A hydrophobic group may contain some hydrophilic groups orsubstituents insofar as the hydrophobic character of the group is notoutweighed. In further variations, a hydrophobic group may includesubstituted silicon atoms, and may include fluorine atoms.

In another instance, hydrophilic groups may be included in the modulesto cause orientation of the modules on surfaces. Examples of hydrophilicgroups include hydroxyl, methoxy, phenol, carboxylic acids and saltsthereof, methyl, ethyl, and vinyl esters of carboxylic acids, amides,amino, cyano, ammonium salts, sulfonium salts, phosphonium salts,polyethylene glycols, epoxy groups, acrylates, sulfonamides, nitro,—OP(O)(OCH₂CH₂N⁺RR′R″)O⁻, guanidinium, aminate, acrylamide, pyridinium,piperidine, and combinations thereof, wherein R, R′ and R″ are eachindependently selected from H or alkyl.

The conformation of a molecule on a surface may depend on the loading,density, or state of the phase or layer in which the molecule resides onthe surface. Surfaces which may be used to orient modules includeinterfaces such as gas-liquid, air-water, immiscible liquid-liquid,liquid-solid, or gas-solid interfaces. The thickness of the orientedlayer may be substantially a monomolecular layer thickness.

A nanofilm is a thin film and may be prepared from macrocyclic modules.A nanofilm may also be prepared from macrocyclic modules in combinationwith other non-modular molecules. In some instances, a nanofilm may beprepared from non-modular molecules. The modules forming a nanofilm maybe deposited on a surface. In some instances, the nanofilm is preparedfrom coupled modules.

A nanofilm composition may be prepared by orienting amphiphilicmacrocyclic modules on a surface. Surface-oriented macrocyclic modulesarranged in a nanofilm layer may provide a unique composition. Thecomposition of a nanofilm prepared from surface-oriented macrocyclicmodules may be solid, gel, or liquid. The modules of the nanofilm may bein an expanded state, a liquid state, or a liquid-expanded state. Thestate of the modules of the nanofilm may be condensed, collapsed, or maybe a solid phase or close-packed state. The modules of the nanofilm mayinteract with each other by weak forces of attraction. The modules of ananofilm prepared from surface-oriented macrocyclic modules need not belinked by any strong interaction or coupling. Alternatively, the modulesof the nanofilm may be linked through, for example, covalent bonds orionic interactions.

As used herein, the term “coupling,” with respect to molecular moietiesor species, molecules, and modules refers to their attachment orassociation with other molecular moieties or species, molecules, ormodules, whether the attachment or association is specific ornon-specific, reversible or non-reversible, is the result of chemicalreaction, or the result of direct or indirect physical interactions,weak interactions, or hydrophobic/hydrophilic interactions, or as theresult of magnetic, electrostatic, or electromagnetic interaction.Coupling may be specific or non-specific, and the bonds formed by acoupling reaction are often covalent bonds, or polar-covalent bonds, ormixed ionic-covalent bonds, and may sometimes be hydrogen bonds, Van derWaals forces, London forces, ionic or electrostatic forces orinteractions, dipole-dipole or dispersive, or other types of binding.

Modules oriented on a surface may be coupled to form a thin layercomposition or nanofilm. Surface-oriented modules may be coupled in atwo-dimensional array to form a substantially monomolecular layernanofilm. The two-dimensional array is generally one molecule thickthroughout the thin layer composition, and may vary locally due tophysical and chemical forces. Coupling of modules may be done to form asubstantially two-dimensional thin film by orienting the modules on asurface before or during the process of coupling.

Macrocyclic modules can be prepared to possess reactive functionalgroups which permit coupling of the modules. The nature of the productsformed by coupling modules depends, in one variation, on the relativeorientations of the reactive functional groups with respect to themodule structure, and in other instances on the arrangement ofcomplementary functional groups on different modules which can formcovalent, non-covalent or other binding attachments with each other.

In one variation, a module includes reactive functional groups whichcouple directly to complementary reactive functional groups of othermodules to form linkages between modules. The reactive functional groupsmay contribute to the amphiphilic character of the module before orafter coupling, and may be covalently or non-covalently attached to themodules. The reactive functional groups may be attached to the modulesbefore, during, or after orientation of the modules on the surface.

Examples of reactive functional groups of modules and the linkagesformed in coupling modules include those shown in Table 2. Each modulemay have 1 to 30 or more reactive functional groups often selected tocouple to another module.

In making nanofilm from macrocyclic modules and other components, one ormore coupling linkages may be formed between macrocyclic modules, andcoupling may occur between macrocyclic modules and other components. Thelinkage formed between macrocyclic modules may be the product of thecoupling of one functional group from each macrocyclic module. Forexample, a hydroxyl group of a first macrocyclic module may couple withan acid group or acid halide group of a second macrocyclic module toform an ester linkage between the two macrocyclic modules. Anotherexample is an imine linkage, —CH═N—, resulting from the reaction of analdehyde, —CH═O, on one macrocyclic module with an amine, —NH₂, onanother macrocyclic module. Examples of linkages between macrocyclicmodules are shown in Table 2.

TABLE 2 Examples of reactive functional groups of modules FunctionalGroup A Functional Group B Linkage Formed —NH₂ —C(O)H —N═CH— —NH₂ —CO₂H—NHC(O)— —NHR —CO₂H —NRC(O)— —OH —CO₂H —OC(O)— —X —O Na —O— —SH —SH—S—S— —X —(NR)Li —NR— —X —S Na —S— —X —NHR —NR— —X —CH₂CuLi —CH₂— —X—(CRR′)_(n=1−6)CuLi —(CRR′)_(n)— module-X module-X module-module —CH₂X—CH₂X —CH₂CH₂— —ONa —C(O)OR —C(O)O— —SNa —C(O)OR —C(O)S— —X —C≡CH —C≡C——C≡CH —C≡CH —C≡C—C≡C— —MgX —C(O)H —CH(OH)— module-NH₂

module-MgX

module-X

—C(O)H —C(O)H —HC═CH— (CH₃)₂C═CH-module module-C(O)Cl

—N═C═O —NH₂ —NHC(O)NH— —N═C═O HO— —NHC(O)O— —C(O)H —NHNH₂ —CH═N—NH— —OH—OC(O)X —OC(O)O— (CH₃)₂C═CH-module module-SH

(CH₃)₂CHC(O)O-module module-CH(O)

module-CH₂C(O)OH module-CH₂C(O)OH

R₂SiH-module

—OP(O)(OH)₂ —OH —OP(O)(OH)O—

In Table 2, R and R′ represent hydrogen or alkyl groups, and X ishalogen or other good leaving group.

These functional groups may be separated from the module by a spacergroup. Examples of spacer groups are alkylene, aryl, acyl, alkoxy,saturated or unsaturated cyclic hydrocarbon, heteroaryl,heteroarylalkyl, or heterocyclic groups, and the correspondingsubstituted groups. Further examples of spacer groups are polymer,copolymer, or oligomer chains, for example, polyethylene oxides,polypropylene oxides, polysaccharides, polylysines, polypeptides,poly(amino acids), polyvinylpyrrolidones, polyesters, polyacrylates,polyamines, polyimines, polystyrenes, poly(vinyl acetate)s,polytetrafluoroethylenes, polyisoprenes, neopropene, polycarbonate,polyvinylchlorides, polyvinylidene fluorides, polyvinylalcohols,polyurethanes, polyamides, polyimides, polysulfones, polyethersulfones,polysulfonamides, polysulfoxides, and copolymers thereof. Examples ofpolymer chain spacer structures include linear, branched, comb anddendrimeric polymers, random and block copolymers, homo-andheteropolymers, flexible and rigid chains. The spacer may be any groupwhich does not interfere with formation of the linkage. A spacer groupmay be substantially longer or shorter than the reactive functionalgroup to which it is attached.

Coupling of surface-oriented modules to each other may occur throughcoupling of reactive functional groups of the modules to linkermolecules. The reactive functional groups involved may be thoseexemplified in Table 2. Modules may couple to at least one other modulethrough a linker molecule. A linker molecule is a discrete molecularspecies used to couple at least two modules. Each module may have 1 to30 or more reactive functional groups which may couple to a linkermolecule. Linker molecules may have 1 to 20 or more reactive functionalgroups which may couple to a module.

In one instance, a linker molecule has at least two reactive functionalgroups, each of which can couple to a module. In these variations,linker molecules may include a variety of reactive functional groups forcoupling modules. Examples of reactive functional groups of modules andlinker molecules are illustrated in Table 3.

TABLE 3 Examples of reactive functional groups of modules and linkermolecules Reactive Reactive Functional Functional Group of Group ofModule A Module B Linker Molecule Linkage —NHR or NH₂ —NHR or NH₂

—NHR or NH₂ —NHR or NH₂

—NHR or NH₂ —NHR or NH₂

—NHR or NH₂ —NHR or NH₂

—OH —OH

—OH —OH

—OH —OH (RO)₂BR′B(OR)₂ —O(HO)BR′B(OH)O— —NHR or NH₂ —NHR or NH₂(RO)₂BR′B(OR)₂ —NH(HO)BR′B(OH)NH— —OH —OH X—(CH₂)_(n)—X —O—(CH₂)_(n)—O——OH —OH ClC(O)—(CH₂)_(n)—C(O)Cl

—NHR or NH₂ —NHR or NH₂

—NHR or NH₂ —NHR or NH₂

In Table 3, n=1-6, m=1-10, R=CH₃ or H, R′=—(CH₂)_(n)— or Phenyl,R″=—(CH₂)—, polyethylene glycol (PEG), or polypropylene glycol (PPG),and X is Br, Cl, I, or other good leaving groups which are organicgroups consisting only of carbon, oxygen, nitrogen, halogen, silicon,phosphorous, sulfur, and hydrogen atoms, and having from 1 to 20 carbonatoms. A module may have a combination of the various reactivefunctional groups exemplified in Table 3.

Methods of initiating coupling of the modules to linker moleculesinclude chemical, thermal, photochemical, electrochemical, andirradiative methods.

In one variation, illustrated in FIG. 1A, module Hexamer 1dh is coupledto a second module through diethyl malonimidate linkers to yield thenanofilm illustrated in FIG. 2B.

A nanofilm comprising interlinked modules can be made by couplingtogether one or more members of the collection of modules, perhaps withother bulky or flexible components, to form a thin layer nanofilmmaterial or composition. Coupling of modules may be complete orincomplete, providing a variety of structural variations useful asnanofilm membranes. The nanofilm may have a unique molecular structure.The compositional structure of a nanofilm made from oriented and coupledmodules may include a unique molecular structure along with othercomponent structures. The structure of the nanofilm may be asubstantially crystalline structure having precise ordering of thecoupled modules over distances which are long compared to the size ofthe modules. Nanofilms may also be prepared having the structure of aglass. Other nanofilms will have less long range ordering of the modulesand be amorphous in molecular form. In some variations, the nanofilm isan elastomeric composition of coupled modules. In other instances, thenanofilm is a brittle thin film. A particular nanofilm may be found tohave regions with more than one of these structural variations.

The thickness of a nanofilm made from surface-oriented molecular speciesor modules is exceptionally small, often being less than about 30nanometers, sometimes less than about 20 nanometers, and sometimes lessthan about 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1. Thethickness of a nanofilm depends partly on the structure and nature ofthe groups on the modules which impart amphiphilic character to themodules. The thickness may be dependent on temperature, and the presenceof solvent on the surface or located within the nanofilm. The thicknessmay be modified if the groups on the modules which impart amphiphiliccharacter to the modules are removed or modified after the modules havebeen coupled, or at other points during or after the process ofpreparation of a nanofilm. The thickness of a nanofilm may also dependon the structure and nature of the surface attachment groups on themodules. The thickness may be modified if the surface attachment groupson the modules are removed or modified after the modules have beencoupled, or at other points during or after the process of preparationof a nanofilm. The thickness of nanofilms made from surface-orientedmolecular species or modules may be less than about 300, 200, 250, 200,150, 100, 90, 80, 70, 60, 50, 40, 30, 20, 10, or 5 Å.

In some instances, the nanofilm may be derivatized to providebiocompatability or reduce fouling of the nanofilm by attachment oradsorption of biomolecules.

The nanofilm composition may include uniquely structured regions inwhich modules are interlinked. Coupling of modules provides a nanofilmin which unique structures may be formed. Nanofilm structures definepores through which atoms, molecules, or particles of only up to acertain size and composition may pass. One variation of a nanofilmstructure includes an area of nanofilm able to face a fluid medium,either liquid or gaseous, and provide pores or openings through whichatoms, ions, small molecules, biomolecules, or other species are able topass. The dimensions of the pores defined by nanofilm structures may beexemplified by quantum mechanical calculations and evaluations, andphysical tests.

The dimensions of the pores defined by nanofilm structures are describedby actual atomic and chemical structural features of the nanofilm. Theapproximate diameters of pores formed in the structure of a nanofilm arefrom about 1-150 Å, or more. In some embodiments, the dimensions of thepores are about 1-10 Å, about 3-15 Å, about 10-15 Å, about 15-20 Å,about 20-30 Å, about 30-40 Å, about 40-50 Å, about 50-75 Å, about 75-100Å, about 100-125 Å, about 125-150 Å, about 150-300 Å, about 300-600 Å,about 600-1000 Å. The approximate dimensions of pores formed in thestructure of a nanofilm are useful to understand the porosity of thenanofilm. On the other hand, the porosity of conventional membranes isnormally quantified by empirical results such as molecular weightcut-off, which reflects complex diffusive and other transportcharacteristics.

In one variation, a nanofilm structure may be an array of coupledmodules which provides an array of pores of substantially uniform size.The pores of uniform size may be defined by the individual modulesthemselves. Each module defines a pore of a particular size, dependingon the conformation and state of the module. For example, theconformation of the interlinked module of the nanofilm may be differentfrom the nascent, pure macrocyclic module in a solvent, and both may bedifferent from the conformation of the amphiphilic module oriented on asurface before coupling. A nanofilm structure including an array ofcoupled modules can provide a matrix or lattice of pores ofsubstantially uniform dimension based on the structure and conformationof the coupled modules.

Modules of various composition and structure may be prepared whichdefine pores of different sizes. A nanofilm prepared from coupledmodules may be made from any one of a variety of modules. Thus,nanofilms having pores of various dimensions are provided, depending onthe particular module used to prepare the nanofilm.

In other instances, nanofilm structures define pores in the matrix ofinterlinked modules. Pores defined by nanofilm structures may have awide range of dimensions, for example, dimensions capable of selectivelyblocking the passage of small molecules or large molecules. Nanofilmstructures may be formed from the coupling of two or more modules, inwhich an interstitial pore is defined by the combined structure of thelinked modules. A nanofilm may have an extended matrix of pores ofvarious dimensions and characteristics. Interstitial pores may be, forexample, less than about 5 Å, less than about 10 Å, about 3-15 Å, about10-15 Å, about 15-20 Å, about 20-30 Å, about 30-40 Å, about 40-50 Å,about 50-75 Å, about 75-100 Å, about 100-125 Å, about 125-150 Å, about150-300 Å, about 300-600 Å, about 600-1000 Å.

The coupling process may result in a nanofilm in which regions of thenanofilm are not precisely monomolecular layers. Various types of localstructures are possible which do not prevent use of the nanofilm in avariety of applications. Local structural features may includeamphiphilic modules which are flipped over relative to neighboringmodules, or turned in a different orientation, having their hydrophobicand hydrophilic facets oriented differently than neighboring modules.Local structural features may also include overlaying of modules inwhich the nanofilm is two or more molecular layers thick, local regionsin which the coupling of the modules is not complete so that some of thecoupling groups of the modules are not coupled to other modules, orlocal regions in which there is an absence of modules. In one variation,the nanofilm has a thickness of up to 30 nanometers due to the layeringof nanofilm structures.

As used herein, a nanofilm comprising “oriented macrocyclic modules”indicates that the macrocyclic modules are substantially uniformlyoriented within the film, but may comprise regions of local structuralfeatures as indicated hereinabove. Local structural features maycomprise, for example, greater than about 30%, less than about 30%, lessthan about 20%, less than about 15%, less than about 10%, less thanabout 5%, less than about 3%, less than about 1% of the surface area ofthe nanofilm. Analogously, a nanofilm comprising “oriented amphiphilicmolecules” indicates that the amphiphilic molecules are substantiallyuniformly oriented within the film, but may comprise regions of localstructural features as indicated hereinabove. Local structural featuresmay comprise, for example, greater than about 30%, less than about 30%,less than about 20%, less than about 15%, less than about 10%, less thanabout 5%, less than about 3%, less than about 1% of the surface area ofthe nanofilm.

A nanofilm may also be prepared with mixtures of different modules, orwith mixtures of macrocyclic modules and other amphiphilic molecules.These nanofilms may have an array of coupled modules and otheramphiphilic molecules in which the positional ordering of the modulesand other amphiphilic molecules is random, or is non-random with regionsin which one type of species is predominant. Nanofilms made frommixtures of different modules, or with mixtures of macrocyclic modulesand other amphiphilic molecules may also have interspersed arrays ofpores of various sizes.

In Langmuir film methods, a monolayer of oriented amphiphilic species isformed on the surface of a liquid subphase. Typically, movable plates orbarriers are used to compress the monolayer and decrease its surfacearea to form a more dense monolayer. At various degrees of compression,having corresponding surface pressures, the monolayer may reach variouscondensed states. In some cases, the film reaches a collapse point atwhich a condensed phase of the monolayer is produced at reduced surfacepressure.

Surface pressure versus film area isotherms are obtained by the Wilhelmybalance method to monitor the state of the film. Extrapolation of theisotherm to zero surface pressure reveals the average surface area permodule before the modules are coupled. When the hydrophobic groups usedto orient the molecules of a Langmuir film monolayer are fatty acidgroups, the collapse typically occurs at a molecular surface area ofabout 20 Å²/molecule. Thus, the isotherm gives empirical indication ofthe state of the thin film.

Nanofilms of oriented species may be deposited on a substrate by variousmethods to provide a porous membrane. For example, description ofLangmuir films and substrates is given in U.S. Pat. Nos. 6,036,778,4,722,856, 4,554,076, and 5,102,798, and in R. A. Hendel et al., Vol.119, J. Am. Chem. Soc. 6909-18 (1997). Description of films onsubstrates is given in Munir Cheryan, Ultrafiltration andMicrofiltration Handbook (1998).

A substrate may be any surface of any material. Substrates may be porousand non-porous. Examples of porous substrates are polymers, track-etchpolycarbonate, track-etch polyester, polyethersulfone, polysulfone,gels, hydrogels, cellulose acetate, polyamide, PVDF, ceramics, anodicalumina, laser ablated polyimide, and UV etched polyacrylate. Examplesof non-porous substrates are silicon, metals, gold, glass, silicates,aluminosilicates, non-porous polymers, and mica.

In forming a nanofilm with Langmuir film methods, a linker molecule,when present, may be added to the solution containing the modules whichis deposited on the surface of the Langmuir subphase. Alternatively, thelinker molecules may be added to the subphase of the Langmuir trough,and subsequently transfer to the module layer phase to couple tomodules. In some instances, modules may be added to the subphase of theLangmuir trough, and subsequently transfer to the module layer phase tocouple to other modules.

Amphiphilic molecules may be oriented on a surface such as an air-waterinterface in a Langmuir trough. The surface oriented amphiphilicmolecules may be compressed to form a Langmuir thin film. Theamphiphilic molecules of the Langmuir thin film may be coupled to eachother or interlinked to form a substantially monomolecular layer thinfilm material. The polar groups of the amphiphilic molecules of theLangmuir thin film may be coupled together by coupling reactions to forma thin film material. The lengths of the hydrophobic tails of theamphiphilic molecules may be from about 8 to 28 carbon atoms. Examplesof hydrophobic tails of the amphiphilic molecules include thehydrophobic groups which may be attached to macrocyclic modules toimpart amphiphilic character to the modules.

Examples of polar groups of the amphiphilic molecules include amide,amino, ester, —SH, acrylate, acrylamide, epoxy, and the hydrophilicgroups as defined above. The polar groups of the amphiphilic moleculesmay be linked directly to each other. For example, sulfhydryl groups maybe coupled to form disulfide links between the amphiphilic molecules ofthe Langmuir thin film. Examples of polar groups include —OH, —OCH₃,—NH₂, —C≡N, —NO₂, —⁺NRR′R″, —SO₃ ⁻, —OPO₂ ²⁻, —OC(O)CH═CH₂, —SO₂NH₂,SO₂NRR′, —OP(O)(OCH₂CH₂N⁺RR′R″)O⁻, —C(O)OH, —C(O)O⁻, guanidinium,aminate, pyridinium, —C(O)OCH₃, —C(O)OCH₂CH₃,

where w is 1-6, —C(O)OCH═CH₂, —O(CH₂)_(x)C(O)NH₂, where x is 1-6,—O(CH₂)_(y)C(O)NHR, where y is 1-6, and —O(CH₂CH₂O)_(z)R, where z is1-6.

The coupling may attach two amphiphilic molecules, such as thealkylthiols of FIG. 2A, by a disulfide link, as illustrated in FIG. 2B.The coupling may attach more than two amphiphilic molecules, forexample, by extended amide linkages. A portion of the amphiphilicmolecules of the nanofilm may be coupled, while the rest are notcoupled. The amphiphilic molecules of the nanofilm, both those which arecoupled and those which are not coupled, may also interact through weaknonbonding or bonding interactions such as hydrogen bonding and otherinteractions.

Pores and barrier properties are found in the structure of the nanofilmmade by coupling amphiphilic molecules. The pores and barrier propertiesmay be modified by the degree or extent of coupling or interaction ofthe amphiphilic molecules, and for example, by the length of the linkermolecules.

Polar groups having ester and amino groups may couple to attach theamphiphilic molecules of FIG. 3A, through amide linkages, to form thenanofilm illustrated in FIG. 3B.

The polar groups of the amphiphilic molecules may be linked to eachother with a linker molecule. For example, amino groups of theamphiphilic molecules of the Langmuir thin film may be coupled byMannich reaction with formaldehyde, as illustrated in FIGS. 4A, 4C and4E. FIGS. 4A, 4C and 4E illustrate a Langmuir film of the amphiphiles,and FIGS. 4B, 4D and 4F illustrate the structures of the linkages formedin the nanofilm upon coupling the amphiphiles. The thin film materialformed by coupling amphiphilic molecules of a Langmuir thin film mayhave barrier properties useful for filtration.

A nanofilm may be prepared from amphiphilic macrocyclic modules orientedon a surface, such as an air-subphase interface in a Langmuir trough,without coupling the modules. Amphiphilic modules may be prepared byattaching groups having interaction with the surface which causeorientation of the modules. A substantially uniform monolayer oforiented amphiphilic modules, such as a Langmuir film, may be formed ona hydrophilic surface. The surface-oriented macrocyclic modules may bearranged in a nanofilm layer, providing a unique composition, which maybe an expanded state, a liquid state, or a liquid-expanded state, or maybe condensed, collapsed, or a solid phase or close-packed state. Thenanofilm of oriented amphiphilic macrocyclic modules may be deposited ona substrate by various methods to provide a porous membrane.

A nanofilm may be prepared by orienting amphiphilic macrocyclic modulesand coupling the modules. Modules may be directly coupled, or may becoupled through linker molecules. Modules may be dissolved in a solventand deposited on an air-subphase interface in a Langmuir trough. Theamphiphilic modules can be oriented at the interface and may becompressed to a condensed thin film.

Linker molecules may be added to the subphase or to the solventcontaining the modules which is deposited on the subphase.

Coupling of modules may be initiated by chemical, thermal,photochemical, electrochemical, and irradiative methods.

The coupling of modules in a nanofilm may attach two or more modules bya linkage or linkages. The coupling may attach more than two modules,for example, by an array of linkages each formed between two modules.Each module may form more than one linkage to another module, and eachmodule may form several types of linkages, including those exemplifiedin Tables 2 and 3. A module may have direct linkages, linkages through alinker molecule, and linkages which include spacers, in any combination.A linkage may connect any portion of a module to any portion of anothermodule. An array of linkages and an array of modules may be described interms of the theory of Bravais lattices and theories of symmetry.

A portion of the modules of a nanofilm may be coupled, while the restare not coupled. The modules of the nanofilm, both those which arecoupled and those which are not coupled, may also interact through weaknon-bonding or bonding interactions such as hydrogen bonding, van derWaals, and other interactions. The arrangement of linkages formed in ananofilm may be represented by a type of symmetry, or may besubstantially unordered.

Functional groups added to the modules to impart amphiphilic characterto the modules may be removed after formation of the nanofilm. Themethod of removal depends on the functional group. For example,lipophilic groups or tails which are attached to modules may beselectively removed. The groups attached to the modules which impartamphiphilic character to the modules may include reactive functionalgroups which can be used to remove the groups at some point during orafter the process of formation of a nanofilm. Acid or base hydrolysismay be used to remove groups attached to the module via a carboxylate oramide linkage. An unsaturated group located in the functional groupwhich imparts amphiphilic character to the module may be oxidized andcleaved by hydrolysis. Photolytic cleavage of the functional group whichimparts amphiphilic character to the module may also be done. Examplesof cleavable functional groups include

where n is zero to four, which is cleavable by light activation, and

where n is zero to four, and m is 7 to 27, which is cleavable by acid orbase catalyzed hydrolysis.

Examples of functional groups added to the modules to impart amphiphiliccharacter to the modules include 8C-28C alkyl groups, —O(8C-28C)alkyl,—NH(8C-28C)alkyl, —OC(O)-(8C-28C)alkyl, —C(O)O-(8C-28C)alkyl,—NHC(O)-(8C-28C)alkyl, —C(O)NH-(8C-28C)alkyl, —CH═CH-(8C-28C)alkyl, and—C≡C-(8C-28C)alkyl, where the carbon atoms of the (8C-28C)alkyl groupsmay be interrupted by one or more —S—, double bond, triple bond or—SiR′R″— groups, substituted with one or more fluorine atoms or anycombination of these, and the R′ and R″ independently comprise(1C-18C)alkyl.

Membranes and Filtration

A membrane is a medium which is brought into contact with a fluid orsolution, separating a species or component from that fluid or solution,for example, for purposes of filtration. Normally, a membrane is asubstance which acts as a barrier to block the passage of some species,while allowing restricted or regulated passage of other species. Ingeneral, permeants may traverse the membrane if they are smaller than acut-off size, or have a molecular weight smaller than a so-calledcut-off molecular weight. The membrane may be called impermeable tospecies which are larger than the cut-off molecular weight. The cut-offsize or molecular weight is a characteristic property of the membrane.Selective permeation is the ability of the membrane to cut-off,restrict, or regulate passage of some species, while allowing smallerspecies to pass. Thus, the selective permeation of a membrane may bedescribed functionally in terms of the largest species able to pass themembrane under given conditions. The size or molecular weight of variousspecies may also be dependent on the conditions in the fluid to beseparated, which may determine the form of the species. For example,species may have a sphere of hydration or solvation in a fluid, and thesize of the species in relation to membrane applications may or may notinclude the water of hydration or the solvent molecules. Thus, amembrane is permeable to a species of a fluid if the species cantraverse the membrane in the form in which it normally would be found inthe fluid. Permeation and permeability may be affected by interactionbetween the species of a fluid and the membrane itself. While varioustheories may describe these interactions, the empirical measurement ofpass/no-pass information relating to a nanofilm, membrane, or module isa useful tool to describe permeation properties. A membrane isimpermeable to a species if the species cannot pass through themembrane.

The nanofilms may have molecular weight species cut offs of, forexample, greater than about 15 kDa, greater than about 10 kDa, greaterthan about 5 kDa, greater that about 1 kDa, greater than about 800 Da,greater than about 600 Da, greater than about 400 Da, greater than about200 Da, greater than about 100 Da, greater than about 50 Da, greaterthan about 20 Da, less than about 15 kDa, less than about 10 kDa, lessthan about 5 kDa, less that about 1 kDa, less than about 800 Da, lessthan about 600 Da, less than about 400 Da, less than about 200 Da, lessthan about 100 Da, less than about 50 Da, less than about 20 Da, about13 kDa, about 190 Da, about 100 Da, about 45 Da, about 20 Da.

“High permeability” indicates a clearance of, for example, greater thanabout 70%, greater than about 80%, greater than about 90% of the solute.“Medium permeability” indicates a clearance of, for example, less thanabout 50%, less than about 60%, less than about 70% of the solute. “Lowpermeability” indicates a clearance of less than, for example, about10%, less than about 20%, less than about 30% of the solute. A membraneis impermeable to a species if it has a very low clearance (for example,less than about 5%, less than about 3%) for the species, or if it hasvery high rejection for the species (for example, greater than about95%, greater than about 98%). The passage or exclusion of a solute ismeasured by its clearance, which reflects the portion of solute thatactually passes through the membrane. For example, the no pass symbol inTables 13-14 indicates that the solute is partly excluded by the module,sometimes less than 90% rejection, often at least 90% rejection,sometimes at least 98% rejection. The pass symbol indicates that thesolute is partly cleared by the module, sometimes less than 90%clearance, often at least 90% clearance, sometimes at least 98%clearance.

A nanofilm may be deposited on a substrate. The deposition may result innon-covalent or weak attachment of the membrane to the substrate throughphysical interactions and weak chemical forces such as van der Waalsforces and weak hydrogen bonding. The membrane may be bound to thesubstrate through ionic or covalent interaction, or other coupling. Ananofilm deposited on a substrate may serve as a membrane. Any number oflayers of nanofilm may be deposited on the substrate to form a membrane.

A layer or layers of various spacing materials may be deposited orattached in between layers of a nanofilm, and a spacing layer may alsobe used in between the substrate and the first deposited layer ofnanofilm. Examples of spacing layer compositions include polymericcompositions, hydrogels (acrylates, poly vinyl alcohols, polyurethanes,silicones), thermoplastic polymers (polyolefins, polyacetals,polycarbonates, polyesters, cellulose esters), polymeric foams,thermosetting polymers, hyperbranched polymers, biodegradable polymerssuch as polylactides, liquid crystalline polymers, polymers made by atomtransfer radical polymerization (ATRP), polymers made by ring openingmetathesis polymerization (ROMP), polyisobutylenes and polyisobutylenestar polymers, and amphiphilic polymers (e.g. Poly(maleic anhydrideoctadecene). Examples of amphiphilic molecules include amphiphilescontaining polymerizable groups such as diynes, enes, or amino-esters.The spacing layers may serve to modify barrier properties of thenanofilm, or may serve to modify transport, flux, or flowcharacteristics of the membrane or nanofilm. Spacing layers may serve tomodify functional characteristics of the membrane or nanofilm, such asstrength, modulus, or other properties.

Deposition of a nanofilm on a substrate may be done byLangmuir-Schaefer, Langmuir-Blodgett, or other methods used withLangmuir systems. In one variation, a nanofilm is deposited on asubstrate in a Langmuir tank by locating the substrate in the subphasebeneath the air-water interface, and lowering the level of the subphaseuntil the nanofilm lands gently on the substrate and is thereforedeposited.

Nanofilms deposited on a substrate may be cured or annealed byradiation, thermal treatment, or drying methods during or afterdeposition on a substrate.

A nanofilm may be attached to a substrate surface by covalent ornon-covalent chemical binding. Surface attachment groups may be providedon the macrocyclic modules which may be used to couple to the substrateto attach the membrane to the substrate. Coupling of some, but not allof the surface attachment groups may be done to attach the nanofilm tothe substrate. Examples of reactive functional groups of modules whichcan be used as surface attachment groups to couple the nanofilm to asubstrate include amine, carboxylic acid, carboxylic ester, alcohol,glycol, vinyl, styrene, epoxide, thiol, magnesium halo or Grignard,acrylate, acrylamide, diene, aldehyde, and mixtures thereof. Table 4illustrates complementary functional groups of the nanofilm and thesubstrate surface which are used to couple the nanofilm to thesubstrate.

TABLE 4 Examples of complementary functional groups of a nanofilm and asubstrate surface indicating linkage formed Nanofilm group Substrategroup Linkage Formed —NH₂ —C(O)H —N═CH— —NH₂ —CO₂H —NHC(O)— —NHR —CO₂H—NRC(O)— —OH —CO₂H —OC(O)— —X —O Na —O— —SH —SH —S—S— —X —(NR)Li —NR— —X—S Na —S— —X —NHR —NR— —X —CH₂CuLi —CH₂— —X —(CRR′)_(n=1−6)CuLi—(CRR′)_(n)— module-X module-X module-module —CH₂X —CH₂X —CH₂CH_(2—)—ONa —C(O)OR —C(O)O— —SNa —C(O)OR —C(O)S— —X —C≡CH —C≡C— —C≡CH —C≡CH—C≡C—C≡C— —MgX —C(O)H —CH(OH)— module-NH₂

module-MgX

module-X

—C(O)H —C(O)H —HC═CH— (CH₃)₂C═CH-module module-C(O)Cl

—N═C═O —NH₂ —NHC(O)NH— —N═C═O HO— —NHC(O)O— —C(O)H —NHNH₂ —CH═N—NH— —OH—OC(O)X —OC(O)O— (CH₃)₂C═CH-module module-SH

(CH₃)₂CHC(O)O-module module-CH(O)

module-CH₂C(O)OH module-CH₂C(O)OH

R₂SiH-module

—OP(O)(OH)₂ —OH —OP(O)(OH)O—

—NH₂

In Table 4, X is halogen or another group leaving group, and R and R′represent hydrogen or alkyl groups. It is to be understood that thefunctional groups listed in Table 4 may be reversed, for example,substrate-NH₂ could couple with nanofilm-C(O)H to form a —N═CH— linkage.It is to be further understood that the functional groups listed inTable 4 may be used in linking modules together, and that the functionalgroups listed in Table 2 may be used to link the nanofilm to thesubstrate.

As illustrated, the substrate in FIG. 5B may have reactive functionalgroups which couple to the reactive functional groups of the modules ofFIG. 5A to attach the membrane to the substrate. The functional groupsof the substrate may be surface groups or linking groups bound to thesubstrate, which may be formed by reactions which bind the surfacegroups or linking groups to the substrate. Surface groups may also becreated on the substrate by a variety of treatments such as cold plasmatreatment, surface etching methods, solid abrasion methods, or chemicaltreatments. Some methods of plasma treatment are given in Inagaki,Plasma Surface Modification and Plasma Polymerization, Technomic,Lancaster, Pa., 1996. In one variation, a photoreactive group such as abenzophenone group is bound to the surface or substrate. Thephotoreactive group may be activated with light, for example,ultraviolet light to provide a reactive species which couples to amacrocyclic module.

The reactive functional groups of the modules and the surface may beblocked with protecting groups until needed. After formation of thecoupled module nanofilm layer, the protecting groups of the reactivefunctional groups which are to be used for attaching the module membranelayer to the substrate surface may be removed, thereby allowing theattachment step to proceed.

Spacer groups may be used to connect reactive functional groups on thenanofilm to the substrate. Spacer groups for surface attachment groupsmay be polymer chains. Examples of polymer chain spacers includepolyethylene oxides, polypropylene oxides, polysaccharides, polylysines,polypeptides, poly(amino acids), polyvinylpyrrolidones, polyesters,polyvinylchlorides, polyvinylidene fluorides, polyvinylalcohols,polyurethanes, polyamides, polyimides, polysulfones, polyethersulfones,polysulfonamides, and polysulfoxides. Examples of polymer chain spacerstructures include linear, branched, comb and dendrimeric polymers,random and block copolymers, homo- and heteropolymers, flexible andrigid chains. Spacer groups for surface attachment groups may alsoinclude bifunctional linker groups or heterobifunctional linker groupsused to couple biomolecules and other chemical species.

Methods of initiating coupling of the modules to the substrate includechemical, thermal, photochemical, electrochemical, and irradiativemethods.

In one instance, a photoreactive group is bound to the substrate. Thephotoreactive group may be activated with light, for example,ultraviolet light to provide a reactive species which couples to ananofilm. The photoreactive species may couple to an atom or group ofthe hydrophilic portion of the nanofilm amphiphiles, or to a hydrophobicportion of the nanofilm amphiphiles. The photoreactive species maycouple to a linkage group of the nanofilm, or other atoms or groupswhich were initially part of the amphiphile or module from which thenanofilm was prepared.

Surface attachment of modules may also be achieved throughligand-receptor mediated interactions, such as biotin-streptavidin. Forexample, the substrate may be coated with streptavidin, and biotin maybe attached to the modules, for example, through linker groups such asPEG or alkyl groups.

Examples of processes in which nanofilms may be useful include processesinvolving liquid or gas as a continuous fluid phase, filtration,clarification, fractionation, pervaporation, reverse osmosis, dialysis,hemodialysis, affinity separation, oxygenation, and other processes.Filtration applications may include ion separation, desalinization, gasseparation, small molecule separation, ultrafiltration, microfiltration,hyperfiltration, water purification, sewage treatment, removal oftoxins, removal of biological species such as bacteria, viruses, orfungus.

Synthons and Macrocyclic Modules

As used herein, the term “alkyl” refers to a branched or unbranchedmonovalent hydrocarbon radical. An “n-mC” alkyl or “(nC-mC)alkyl” refersto all alkyl groups containing from n to m carbon atoms. For example, a1-4C alkyl refers to a methyl, ethyl, propyl, or butyl group. Allpossible isomers of an indicated alkyl are also included. Thus, propylincludes isopropyl, butyl includes n-butyl, isobutyl and t-butyl, and soon. An alkyl group with from 1-6 carbon atoms is referred to as “loweralkyl.” The term alkyl includes substituted alkyls. As used herein, theterm “substituted alkyl” refers to an alkyl group with an additionalgroup or groups attached to any carbon of the alkyl group. Additionalgroups may include one or more functional groups such as alkyl, loweralkyl, aryl, acyl, halogen, alkylhalo, hydroxy, amino, alkoxy,alkylamino, acylamino, acyloxy, aryloxy, aryloxyalkyl, mercapto, bothsaturated and unsaturated cyclic hydrocarbons, heterocycles, and others.

As used herein, the term “alkenyl” refers to any structure or moietyhaving the unsaturation C═C. As used herein, the term “alkynyl” refersto any structure or moiety having the unsaturation C≡C.

As used herein, the terms “R,” “R′,” “R″”, and “R′″” in a chemicalformula refer to a hydrogen or a functional group, each independentlyselected, unless stated otherwise.

As used herein, the term “aryl” refers to an aromatic group which may bea single aromatic ring or multiple aromatic rings which are fusedtogether, linked covalently, or linked to a common group such as amethylene, ethylene, or carbonyl, and includes polynuclear ringstructures. An aromatic ring or rings may include substituted orunsubstituted phenyl, naphthyl, biphenyl, diphenylmethyl, andbenzophenone groups, among others. The term “aryl” includes substitutedaryls.

As used herein, the term “substituted aryl” refers to an aryl group withan additional group or groups attached to any carbon of the aryl group.Additional groups may include one or more functional groups such aslower alkyl, aryl, acyl, halogen, alkylhalos, hydroxy, amino, alkoxy,alkylamino, acylamino, acyloxy, aryloxy, aryloxyalkyl, thioether,heterocycles, both saturated and unsaturated cyclic hydrocarbons whichare fused to the aromatic ring(s), linked covalently or linked to acommon group such as a methylene or ethylene group, or a carbonyllinking group such as in cyclohexyl phenyl ketone, and others.

As used herein, the term “heteroaryl” refers to an aromatic ring(s) inwhich one or more carbon atoms of the aromatic ring(s) are substitutedby a heteroatom such as nitrogen, oxygen, or sulfur. Heteroaryl refersto structures which may include a single aromatic ring, multiplearomatic rings, or one or more aromatic rings coupled to one or morenonaromatic rings. It includes structures having multiple rings, fusedor unfused, linked covalently, or linked to a common group such as amethylene or ethylene group, or linked to a carbonyl as in phenylpyridyl ketone. As used herein, the term “heteroaryl” includes ringssuch as thiophene, pyridine, isoxazole, phthalimide, pyrazole, indole,furan, or benzo-fused analogues of these rings.

As used herein, the term “acyl” refers to a carbonyl substituent,—C(O)R, where R is alkyl or substituted alkyl, aryl or substituted aryl,which may be called an alkanoyl substituent when R is alkyl.

As used herein, the term “amino” refers to a group —NRR′, where R and R′may independently be hydrogen, lower alkyl, substituted lower alkyl,aryl, substituted aryl or acyl.

As used herein, the term “alkoxy” refers to an —OR group, where R is analkyl, substituted lower alkyl, aryl, substituted aryl. Alkoxy groupsinclude, for example, methoxy, ethoxy, phenoxy, substituted phenoxy,benzyloxy, phenethyloxy, t-butoxy, and others.

As used herein, the term “thioether” refers to the general structureR—S—R′ in which R and R′ are the same or different and may be alkyl,aryl or heterocyclic groups. The group —SH may also be referred to as“sulfhydryl” or “thiol” or “mercapto.”

As used herein, the term “saturated cyclic hydrocarbon” refers to ringstructures cyclopropyl, cyclobutyl, cyclopentyl groups, and others,including substituted groups. Substituents to saturated cyclichydrocarbons include substituting one or more carbon atoms of the ringwith a heteroatom such as nitrogen, oxygen, or sulfur. Saturated cyclichydrocarbons include bicyclic structures such as bicycloheptanes andbicyclooctanes, and multicyclic structures.

As used herein, the term “unsaturated cyclic hydrocarbon” refers to amonovalent nonaromatic group with at least one double bond, such ascyclopentene, cyclohexene, and others, including substituted groups.Substituents to unsaturated cyclic hydrocarbons include substituting oneor more carbon atoms of the ring with a heteroatom such as nitrogen,oxygen, or sulfur. As used herein, the term “cyclic hydrocarbon”includes substituted and unsubstituted, saturated and unsaturated cyclichydrocarbons, and multicyclic structures. Unsaturated cyclichydrocarbons include bicyclic structures such as bicycloheptenes andbicyclooctenes, and multicyclic structures.

As used herein, the term “heteroarylalkyl” refers to alkyl groups inwhich the heteroaryl group is attached through an alkyl group.

As used herein, the term “heterocyclic” refers to a monovalent saturatedor unsaturated nonaromatic group having a single ring or multiplecondensed rings from 1-12 carbon atoms and from 1-4 heteroatoms selectedfrom nitrogen, phosphorous, sulfur, or oxygen within the ring. Examplesof heterocycles include tetrahydrofuran, morpholine, piperidine,pyrrolidine, and others.

As used herein, each chemical term described above expressly includesthe corresponding substituted group. For example, the term“heterocyclic” includes substituted heterocyclic groups.

As used herein, the terms “amphiphile” or “amphiphilic” refer to aspecies which exhibits both hydrophilic and lipophilic character. Ingeneral, an amphiphile contains a lipophilic moiety and a hydrophilicmoiety. An amphiphile may form a Langmuir film.

Examples of hydrophilic moieties include, without limitation, hydroxyl,methoxy, phenol, carboxylic acids and salts thereof, methyl and ethylesters of carboxylic acids, amides, amino, cyano, ammonium salts,monoalkyl-substituted amino groups, di-alkyl-substituted amino groups,—NRR′, —N≡C, —NHR, sulfonium salts, phosphonium salts,polyethyleneglycols, polypropyleneglycols, epoxy groups, acrylates,sulfonamides, nitro, —OP(O)(OCH₂CH₂N⁺RR′R″)O⁻, guanidinium, aminate,acrylamide, and pyridinium. Such hydrophilic moieties may include groupssuch as polyethylene glycols, or for example, polymethylene chainssubstituted with alcohol, carboxylate, acrylate, methacrylate, or

groups, where y is 1-6. Hydrophilic moieties may also include alkylchains having internal amino or substituted amino groups, for example,internal —NH—, —NC(O)R—, or —NC(O)CH═CH₂— groups. Hydrophilic moietiesmay also include polycaprolactones, polycaprolactone diols, poly(aceticacid)s, poly(vinyl acetates)s, poly(2-vinyl pyridine)s, celluloseesters, cellulose hydroxyl ethers, poly(L-lysine hydrobromide)s,poly(itaconic acid)s, poly(maleic acid)s, poly(styrenesulfonic acid)s,poly(aniline)s, or poly(vinyl phosphonic acid)s.

Examples of lipophilic moieties include, without limitation, linear orbranched alkyls, including 1-28C hydrocarbons. Examples of groups whichmay be coupled to a synthon or macrocyclic module as a lipophilic groupinclude alkyls, —CH═CH—R, —C≡C—R, —OC(O)—R, —C(O)O—R, —NHC(O)—R,—C(O)NH—R, and —O—R, where R is 4-18C alkyl. Each chain mayindependently comprise, without limitation, alkenyl, alkynyl, saturatedand unsaturated cyclic hydrocarbons, or aromatic groups. Each chain mayalso contain, interspersed among the carbons of the chain, one or moresilicon atoms substituted with alkyl, alkenyl, alkynyl, saturated andunsaturated cyclic hydrocarbons, or aryl groups. The carbon atoms ofeach chain may independently be substituted with one or more fluorineatoms. The carbon atoms of an alkyl group may be interrupted by one ormore functional groups such as, for example, —S—, double bond, triplebond or —SiR′R″— groups (where R′ and R″ are independently H or alkyl),any of which may be substituted with one or more fluorine atoms, and anycombination of such functional groups may be used.

As used herein, the term “functional group” includes, but is not limitedto, chemical groups, organic groups, inorganic groups, organometallicgroups, aryl groups, heteroaryl groups, cyclic hydrocarbon groups, amino(—NH₂), hydroxyl (—OH), cyano (—C≡N), nitro (NO₂), carboxyl (—COOH),formyl (—CHO), keto (—CH₂C(O)CH₂—), alkenyl (—C═C—), alkynyl, (—C≡C—),and halo (F, Cl, Br and I) groups.

As used herein, the term “activated acid” refers to a —C(O)X moiety,where X is a leaving group, in which the X group is readily displaced bya nucleophile to form a covalent bond between the —C(O)— and thenucleophile. Examples of activated acids include acid chlorides, acidfluorides, p-nitrophenyl esters, pentafluorophenyl esters, andN-hydroxysuccinimide esters.

As used herein, the term “amino acid residue” refers to the productformed when a species comprising at least one amino (—NH₂) and at leastone carboxyl (—C(O)O—) group couples through either of its amino orcarboxyl groups with an atom or functional group of a synthon. Whicheverof the amino or carboxyl groups is not involved in the coupling may beblocked with a removable protective group.

Synthons

As used herein, the term “synthon” refers to a molecule used to make amacrocyclic module. A synthon may be substantially one isomericconfiguration, for example, a single enantiomer. A synthon may besubstituted with functional groups which are used to couple a synthon toanother synthon or synthons, and which are part of the synthon. Asynthon may be substituted with an atom or group of atoms which are usedto impart hydrophilic, lipophilic, or amphiphilic character to thesynthon or to species made from the synthon. A synthon may besubstituted with an atom or group of atoms to form one or morefunctional groups on the synthon which may be used to couple the synthonto another synthon or synthons. The synthon before being substitutedwith functional groups or groups used to impart hydrophilic, lipophilic,or amphiphilic character may be called the core synthon. As used herein,the term “synthon” refers to a core synthon, and also refers to asynthon substituted with functional groups or groups used to imparthydrophilic, lipophilic, or amphiphilic character.

As used herein, the term “cyclic synthon” refers to a synthon having oneor more ring structures. Examples of ring structures include aryl,heteroaryl, and cyclic hydrocarbon structures including bicyclic ringstructures and multicyclic ring structures. Examples of core cyclicsynthons include, but are not limited to, benzene, cyclohexadiene,cyclopentadiene, naphthalene, anthracene, phenylene, phenanthracene,pyrene, triphenylene, phenanthrene, pyridine, pyrimidine, pyridazine,biphenyl, bipyridyl, cyclohexane, cyclohexene, decalin, piperidine,pyrrolidine, morpholine, piperazine, pyrazolidine, quinuclidine,tetrahydropyran, dioxane, tetrahydrothiophene, tetrahydrofuran, pyrrole,cyclopentane, cyclopentene, triptycene, adamantane,bicyclo[2.2.1]heptane, bicyclo[2.2.1]heptene, bicyclo[2.2.2]octane,bicyclo[2.2.2]octene, bicyclo[3.3.0]octane, bicyclo[3.3.0]octene,bicyclo[3.3.1]nonane, bicyclo[3.3.1]nonene, bicyclo[3.2.2]nonane,bicyclo[3.2.2]nonene, bicyclo[4.2.2]decane, 7-azabicyclo[2.2.1]heptane,1,3-diazabicyclo[2.2.1]heptane, and spiro[4.4]nonane. A core synthoncomprises all isomers or arrangements of coupling the core synthon toother synthons. For example, the core synthon benzene includes synthonssuch as 1,2- and 1,3-substituted benzenes, where the linkages betweensynthons are formed at the 1,2- and 1,3-positions of the benzene ring,respectively. For example, the core synthon benzene includes1,3-substituted synthons such as

where L is a linkage between synthons and the 2,4,5,6 positions of thebenzene ring may also have substituents. A condensed linkage betweensynthons involves a direct coupling between a ring atom of one cyclicsynthon to a ring atom of another cyclic synthon, for example, wheresynthons M-X and M-X couple to form M-M, where M is a cyclic synthon andX is halogen; as for example when M is phenyl resulting in the condensedlinkage

Macrocyclic Modules

A macrocyclic module is a closed ring of coupled synthons. To make amacrocyclic module, synthons may be substituted with functional groupsto couple the synthons to form a macrocyclic module. Synthons may alsobe substituted with functional groups which will remain in the structureof the macrocyclic module. Functional groups which remain in themacrocyclic module may be used to couple the macrocyclic module to othermacrocyclic modules.

A macrocyclic module may contain from three to about twenty-four cyclicsynthons. In the closed ring of a macrocyclic module, a first cyclicsynthon may be coupled to a second cyclic synthon, the second cyclicsynthon may be coupled to a third cyclic synthon, the third cyclicsynthon may be coupled to a fourth cyclic synthon, if four cyclicsynthons are present in the macrocyclic module, the fourth to a fifth,and so on, until an n^(th) cyclic synthon may be coupled to itspredecessor, and the n^(th) cyclic synthon may be coupled to the firstcyclic synthon to form a closed ring of cyclic synthons. In onevariation, the closed ring of the macrocyclic module may be formed witha linker molecule.

A macrocyclic module may be an amphiphilic macrocyclic module whenhydrophilic and lipophilic functional groups exist in the structure. Theamphiphilic character of a macrocyclic module may arise from atoms inthe synthons, in the linkages between synthons, or in functional groupscoupled to the synthons or linkages.

In some variations, one or more of the synthons of a macrocyclic modulemay be substituted with one or more lipophilic moieties, while one ormore of the synthons may be substituted with one or more hydrophilicmoieties, thereby forming an amphiphilic macrocyclic module. Lipophilicand hydrophilic moieties may be coupled to the same synthon or linkagein an amphiphilic macrocyclic module. Lipophilic and hydrophilicmoieties may be coupled to the macrocyclic module before or afterformation of the closed ring of the macrocyclic module. For example,lipophilic or hydrophilic moieties may be added to the macrocyclicmodule after formation of the closed ring by substitution of a synthonor linkage.

The amphiphilicity of a macrocyclic module may be characterized in partby its ability to form a stable Langmuir film. A Langmuir film may beformed on a Langmuir trough at a particular surface pressure measured inmilliNewtons per meter (mN/m) with a particular barrier speed measuredin millimeters per minute (mm/min), and the isobaric creep or change infilm area at constant surface pressure can be measured to characterizestability of the film. For example, a stable Langmuir film ofmacrocyclic modules on a water subphase may have an isobaric creep at5-15 mN/m such that the majority of the film area is retained over aperiod of time of about one hour. Examples of stable Langmuir films ofmacrocyclic modules on a water subphase may have isobaric creep at 5-15mN/m such that about 70% of the film area is retained over a period oftime of about 30 minutes, sometimes about 70% of the film area isretained over a period of time of about 40 minutes, sometimes about 70%of the film area is retained over a period of time of about 60 minutes,and sometimes about 70% of the film area is retained over a period oftime of about 120 minutes. Other examples of stable Langmuir films ofmacrocyclic modules on a water subphase may have isobaric creep at 5-15mN/m such that about 80% of the film area is retained over a period oftime of about thirty minutes, sometimes about 85% of the film area isretained over a period of time of about thirty minutes, sometimes about90% of the film area is retained over a period of time of about thirtyminutes, sometimes about 95% of the film area is retained over a periodof time of about thirty minutes, and sometimes about 98% of the filmarea is retained over a period of time of about thirty minutes.

In one aspect, an individual macrocyclic module may include a pore inits structure. Each macrocyclic module may define a pore of a particularsize, depending on the conformation and state of the module. Variousmacrocyclic modules may be prepared which define pores of differentsizes.

A macrocyclic module may include a flexibility in its structure.Flexibility may permit a macrocyclic module to more easily form linkageswith other macrocyclic modules by coupling reactions. Flexibility of amacrocyclic module may also play a role in regulating passage of speciesthrough the pore of the macrocyclic module. For example, flexibility mayaffect the dimension of the pore of an individual macrocyclic modulesince various conformations may be available to the structure. Forexample, the macrocyclic module may have a certain pore dimension in oneconformation when no substituents are located at the pore, and the samemacrocyclic module may have a different pore dimension in anotherconformation when one or more substituents of that macrocycle arelocated at the pore. Likewise, a macrocyclic module may have a certainpore dimension in one conformation when one group of substituents arelocated at the pore, and have a different pore dimension in a differentconformation when a different group of substituents are located at thepore. For example, the “one group” of substituents located at the poremay be three alkoxy groups arranged in one regioisomer, while the“different group” of substituents may be two alkoxy groups arranged inanother regioisomer. The effect of the “one group” of substituentslocated at the pore and the “different group” of substituents located atthe pore is to provide a macrocyclic module composition which mayregulate transport and filtration, in conjunction with other regulatingfactors.

In making macrocyclic modules from synthons, the synthons may be used asa substantially pure single isomer, for example, as a pure singleenantiomer.

In making macrocyclic modules from synthons, one or more couplinglinkages are formed between adjacent synthons. The linkage formedbetween synthons may be the product of the coupling of one functionalgroup on one synthon to a complementary functional group on a secondsynthon. For example, a hydroxyl group of a first synthon may couplewith an acid group or acid halide group of a second synthon to form anester linkage between the two synthons. Another example is an iminelinkage, —CH═N—, resulting from the reaction of an aldehyde, —CH═O, onone synthon with an amine, —NH₂, on another synthon. Examples ofcomplementary functional groups and linkages between synthons are shownin Table 5.

TABLE 5 EXAMPLES OF FUNCTIONAL GROUPS OF SYNTHONS AND SYNTHON LINKAGESFunctional Group A Functional Group B Linkage Formed —NH₂ —C(O)H —N═CH——NH₂ —CO₂H —NHC(O)— —NHR —CO₂H —NRC(O)— —OH —CO₂H —OC(O)— —X —O Na —O——SH —SH —S—S— —X —(NR)Li —NR— —X —S Na —S— —X —NHR —NR— —X —CH₂CuLi—CH₂— —X —(CRR′)_(n=1−6)CuLi —(CRR′)_(n)— synthon-X synthon-Xsynthon-synthon —CH₂X —CH₂X —CH₂CH₂— —ONa —C(O)OR —C(O)O— —SNa —C(O)OR—C(O)S— —X —C≡CH —C≡C— —C≡CH —C≡CH —C≡C—C≡C— —MgX —C(O)H —CH(OH)—synthon-NH₂

synthon-MgX

synthon-X

—C(O)H —C(O)H —HC═CH— (CH₃)₂C═CH-synthon synthon-C(O)Cl

—N═C═O —NH₂ —NHC(O)NH— —N═C═O HO— —NHC(O)O— —C(O)H —NHNH₂ —CH═N—NH— —OH—OC(O)X —OC(O)O— (CH₃)₂C═CH-synthon synthon-SH

(CH₃)₂CHC(O)O— synthon synthon-CH(O)

synthon-CH₂C(O)OH synthon-CH₂C(O)OH

R₂SiH-synthon

—OP(O)(OH)₂ —OH —OP(O)(OH)O—

In Table 5, R and R′ represent hydrogen or a functional group, and X ishalogen or other good leaving groups.

In another variation, a macrocyclic module may have functional groupsfor coupling to other macrocyclic modules which were coupled to themacrocyclic module after initial preparation of the closed ring. Forexample, an imine linkage of a macrocyclic module may be substitutedwith one of various functional groups to produce additional macrocyclicmodules. Examples of linkages between synthons having functional groupsfor coupling macrocyclic modules are shown in Table 6.

TABLE 6 EXAMPLES OF FUNCTIONAL GROUPS OF SYNTHONS AND SYNTHON LINKAGESFunctional Group of Macrocyclic Module Linkage Reagent SubstitutedLinkage

In Table 6, X is halogen, and Q₁ and Q₂ are independently selectedsynthons which are part of a module.

The functional groups of synthons used to form linkages between synthonsor other macrocyclic modules may be separated from the synthon by aspacer. A spacer can be any atom or group of atoms which couples thefunctional group to the synthon, and does not interfere with thelinkage-forming reaction. A spacer is part of the functional group, andbecomes part of the linkage between synthons. An example of a spacer isa methylene group, —CH₂—. The spacer may be said to extend the linkagebetween synthons. For example, if one methylene spacer were inserted inan imine linkage, —CH═N—, the resulting imine linkage may be —CH₂CH—N—.

A linkage between synthons may also contain one or more atoms providedby an external moiety other than the two functional groups of thesynthons. An external moiety may be a linker molecule which may couplewith the functional group of one synthon to form an intermediate whichcouples with a functional group on another synthon to form a linkagebetween the synthons, such as, for example, to form a closed ring ofsynthons from a series of coupled synthons. An example of a linkermolecule is formaldehyde. For example, amino groups on two synthons mayundergo Mannich reaction in the presence of formaldehyde as the linkermolecule to produce the linkage —NHCH₂NH—.

The macrocyclic module compositions may comprise, for example, fromthree to about twenty-four cyclic synthons coupled to form a closedring; at least two functional groups for coupling the closed ring tocomplementary functional groups on at least two other closed rings;wherein each functional group and each complementary functional groupcomprises a functional group containing atoms selected from the groupconsisting of C, H, N, O, Si, P, S, B, Al, halogens, and metals from thealkali and alkaline earth groups. In some embodiments, the macrocyclicmodule may comprise at least two closed rings coupled through saidfunctional groups. In another embodiment, the macrocyclic modulecomprises at least three closed rings coupled through said functionalgroups.

In another embodiment, the macrocyclic modules may comprise from threeto about twenty-four cyclic synthons coupled to form a closed ringdefining a pore; the closed ring having a first pore dimension in afirst conformation when a first group of substituents is located at thepore and a second pore dimension in a second conformation when a secondgroup of substituents is located at the pore;

wherein each substituent of each group comprises a functional groupcontaining atoms selected from the group consisting of C, H, N, O, Si,P, S, B, Al, halogens, and metals from the alkali and alkaline earthgroups.

In another example, the macrocyclic module comprises (a) from three toabout twenty-four cyclic synthons coupled to form a closed ring defininga pore; (b) at least one functional group coupled to the closed ring atthe pore and selected to transport a selected species through the pore,wherein the at least one functional group comprises a functional groupcontaining atoms selected from the group consisting of C, H, N, O, Si,P, S, B, Al, halogens, and metals from the alkali and alkaline earthgroups; (c) a selected species to be transported through the pore. Insome embodiments, the selected species is selected from the group ofovalbumin, glucose, creatinine, H₂PO₄ ⁻, HPO₄ ⁻², HCO₃ ⁻, urea, Na⁺,Li⁺, and K⁺.

In some embodiments, the macrocyclic module composition is coupled to asolid support selected from the group of Wang resins, hydrogels,aluminas, metals, ceramics, polymers, silica gels, sepharose, sephadex,agarose, inorganic solids, semiconductors, and silicon wafers. In otherembodiments, the macrocyclic module composition retains at least 85% offilm area after thirty minutes on a Langmuir trough at 5-15 mN/m. Inother embodiments, the macrocyclic module composition retains at least95% of film area after thirty minutes on a Langmuir trough at 5-15 mN/m.In another embodiment, the macrocyclic module retains at least 98% offilm area after thirty minutes on a Langmuir trough at 5-15 mN/m.

In some embodiments, the cyclic synthons are each independently selectedfrom the group consisting of benzene, cyclohexadiene, cyclohexene,cyclohexane, cyclopentadiene, cyclopentene, cyclopentane, cycloheptane,cycloheptene, cycloheptadiene, cycloheptatriene, cyclooctane,cyclooctene, cyclooctadiene, cyclooctatriene, cyclooctatetraene,naphthalene, anthracene, phenylene, phenanthracene, pyrene,triphenylene, phenanthrene, pyridine, pyrimidine, pyridazine, biphenyl,bipyridyl, decalin, piperidine, pyrrolidine, morpholine, piperazine,pyrazolidine, quinuclidine, tetrahydropyran, dioxane,tetrahydrothiophene, tetrahydrofuran, pyrrole, triptycene, adamantane,bicyclo[2.2.1]heptane, bicyclo[2.2.1]heptene, bicyclo[2.2.2]octane,bicyclo[2.2.2]octene, bicyclo[3.3.0]octane, bicyclo[3.3.0]octene,bicyclo[3.3.1]nonane, bicyclo[3.3.1]nonene, bicyclo[3.2.2]nonane,bicyclo[3.2.2]nonene, bicyclo[4.2.2]decane, 7-azabicyclo[2.2.1]heptane,1,3-diazabicyclo[2.2.1]heptane, and spiro[4.4]nonane.

In other embodiments, each coupled cyclic synthon is independentlycoupled to two adjacent synthons by a linkage selected from the groupconsisting of (a) a condensed linkage, and (b) a linkage selected fromthe group consisting of —NRC(O)—, —OC(O)—, —O—, —S—S—, —S—, —NR—,—(CRR′)_(p)—, —CH₂NH—, —C(O)S—, —C(O)O—, —C≡C—, —C≡C—C≡C—, —CH(OH)—,—HC═CH—, —NHC(O)NH—, —NHC(O)O—, —NHCH₂NH—, —NHCH₂CH(OH)CH₂NH—,—N═CH(CH₂)_(p)CH═N—, —CH₂CH(OH)CH₂—, —N═CH(CH₂)_(h)CH═N— where h is 1-4,—CH═N—NH—, —OC(O)O—, —OP(O)(OH)O—, —CH(OH)CH₂NH—, —CH(OH)CH₂—,—CH(OH)C(CH₃)₂C(O)O—,

where G is halogen,

wherein p is 1-6;wherein R and R′ are each independently selected from the group ofhydrogen and alkyl; wherein the linkage is independently configured ineither of two possible configurations, forward and reverse, with respectto the synthons it couples together, if the two configurations aredifferent structures; wherein Q is one of the synthons connected by thelinkage.

In other embodiment, a closed ring composition may be comprised of theformula:

wherein: J is 2-23; Q¹ are synthons each independently selected from thegroup consisting of (a) phenyl synthons coupled to linkages L at1,2-phenyl positions, (b) phenyl synthons coupled to linkages L at1,3-phenyl positions, (c) aryl synthons other than phenyl synthons, (d)heteroaryl synthons other than pyridinium synthons, (e) saturated cyclichydrocarbon synthons, (f) unsaturated cyclic hydrocarbon synthons, (g)saturated bicyclic hydrocarbon synthons, (h) unsaturated bicyclichydrocarbon synthons, (i) saturated multicyclic hydrocarbon synthons,and (j) unsaturated multicyclic hydrocarbon synthons; wherein ringpositions of each Q¹ which are not coupled to a linkage L aresubstituted with hydrogen or functional groups containing atoms selectedfrom the group of C, H, N, O, Si, P, S, B, Al, halogens, and metals fromthe alkali and alkaline earth groups; Q² is a synthon independentlyselected from the group consisting of (a) aryl synthons other thanphenyl synthons and naphthalene synthons coupled to linkages L at2,7-naphthyl positions, (b) heteroaryl synthons other than pyridinesynthons coupled to linkages L at 2,6-pyridino positions, (c) saturatedcyclic hydrocarbon synthons other than cyclohexane synthons coupled tolinkages L at 1,2-cyclohexyl positions, (d) unsaturated cyclichydrocarbon synthons other than pyrrole synthons coupled to linkages Lat 2,5-pyrrole positions, (e) saturated bicyclic hydrocarbon synthons,(f) unsaturated bicyclic hydrocarbon synthons, (g) saturated multicyclichydrocarbon synthons, and (h) unsaturated multicyclic hydrocarbonsynthons; wherein ring positions of Q² which are not coupled to an L aresubstituted with hydrogen or functional groups containing atoms selectedfrom the group consisting of C, H, N, O, Si, P, S, B, Al, halogens, andmetals from the alkali and alkaline earth groups; L are linkages betweenthe synthons each independently selected from the group consisting of(a) a condensed linkage, and (b) a linkage selected from the groupconsisting of —NRC(O)—, —OC(O)—, —O—, —S—S—, —S—, —NR—, —(CRR′)_(p)—,—CH₂NH—, —C(O)S—, —C(O)O—, —C≡C—, —C≡C—C≡C—, —CH(OH)—, —HC═CH—,—NHC(O)NH—, —NHC(O)O—, —NHCH₂NH—, —NHCH₂CH(OH)CH₂NH—,—N═CH(CH₂)_(p)CH═N—, —CH₂CH(OH)CH₂—, —N═CH(CH₂)_(h)CH═N— where h is 1-4,—CH═N—NH—, —OC(O)O—, —OP(O)(OH)O—, —CH(OH)CH₂NH—, —CH(OH)CH₂—,—CH(OH)C(CH₃)₂C(O)O—,

where G is halogen,

wherein p is 1-6; wherein R and R′ are each independently selected fromthe group of hydrogen and alkyl; wherein linkages L are eachindependently configured in either of two possible configurations,forward and reverse, with respect to the synthons it couples together,if the two configurations are different structures; wherein y is 1 or 2,and Q^(y) are each independently one of the Q¹ or Q² synthons connectedby the linkage.

In some embodiments, the functional groups are each independentlyselected from the group consisting of hydrogen, an activated acid, —OH,—C(O)OH, —C(O)H, —C(O)OCH₃, —C(O)Cl, —NRR, —NRR⁺, —MgX, —Li, —OLi, —OK,—ONa, —SH, —C(O)(CH₂)₂C(O)OCH₃, —NH-alkyl-C(O)CH₂CH(NH₂)CO₂-alkyl,—CH═CH₂, —CH═CHR, —CH═CR₂, 4-vinylaryl, —C(O)CH═CH₂, —NHC(O)CH═CH₂,—C(O)CH═CH(C₆H₅),

—OH, —OC(O)(CH₂)₂C(O)OCH₃, —OC(O)CH═CH₂,

and

—P(O)(OH)(OX), —P(═O)(O⁻)O(CH₂)_(s)NR₃ ⁺; wherein R are eachindependently selected from the group consisting of hydrogen and 1-6Calkyl; X is selected from the group consisting of Cl, Br, and I; r is1-50; and s is 1-4.

In another example, a closed ring composition comprises the formula:

wherein: J is 2-23; Q¹ are synthons each independently selected from thegroup consisting of (a) phenyl synthons coupled to linkages L at1,2-phenyl positions, (b) phenyl synthons coupled to linkages L at1,3-phenyl positions, and (c) cyclohexane synthons coupled to linkages Lat 1,2-cyclohexyl positions; wherein ring positions of each Q¹ which arenot coupled to a linkage L are substituted with hydrogen or functionalgroups containing atoms selected from the group of C, H, N, O, Si, P, S,B, Al, halogens, and metals from the alkali and alkaline earth groups;Q² is a cyclohexane synthon coupled to linkages L at 1,2-cyclohexylpositions; wherein ring positions of Q² which are not coupled to an Lare substituted with functional groups containing atoms selected fromthe group consisting of C, H, N, O, Si, P, S, B, Al, halogens, andmetals from the alkali and alkaline earth groups; L are linkages betweenthe synthons each independently selected from the group consisting of(a) a condensed linkage, and (b) a linkage selected from the groupconsisting of —NRC(O)—, —OC(O)—, —O—, —S—S—, —S—, —NR—, —(CRR′)_(p)—,—CH₂NH—, —C(O)S—, —C(O)O—, —C≡C—, —C≡C—C≡C—, —CH(OH)—, —HC═CH—,—NHC(O)NH—, —NHC(O)O—, —NHCH₂NH—, —NHCH₂CH(OH)CH₂NH—,—N═CH(CH₂)_(p)CH═N—, —CH₂CH(OH)CH₂—, —N═CH(CH₂)_(h)CH═N— where h is 1-4,—CH═N—NH—, —OC(O)O—, —OP(O)(OH)O—, —CH(OH)CH₂NH—, —CH(OH)CH₂—,—CH(OH)C(CH₃)₂C(O)O—,

where G is halogen,

wherein p is 1-6; wherein R and R′ are each independently selected fromthe group of hydrogen and alkyl; wherein linkages L are eachindependently configured in either of two possible configurations,forward and reverse, with respect to the synthons it couples together,if the two configurations are different structures; wherein y is 1 or 2,and Q^(y) are each independently one of the Q¹ or Q² synthons connectedby the linkage.

In some cases, the functional groups are each independently selectedfrom the group consisting of hydrogen, an activated acid, —OH, —C(O)OH,—C(O)H, —C(O)OCH₃, —C(O)Cl, —NRR, —NRRR⁺, —MgX, —Li, —OLi, —OK, —ONa,—SH, —C(O)(CH₂)₂C(O)OCH₃, —NH-alkyl-C(O)CH₂CH(NH₂)CO-alkyl, —CH═CH₂,—CH═CHR, —CH═CR₂, 4-vinylaryl, —C(O)CH═CH₂, —NHC(O)CH═CH₂,—C(O)CH═CH(C₆H₅),

—OH, —OC(O)(CH₂)₂C(O)OCH₃, —OC(O)CH═CH₂,

and

—P(O)(OH)(OX), —P(═O)(O⁻)O(CH₂)_(s)NR₃ ⁺;wherein R are each independently selected from the group consisting ofhydrogen and 1-6C alkyl; X is selected from the group consisting of Cl,Br, and I; r is 1-50; and s is 1-4.

In another embodiment is a closed ring composition of the formula:

wherein: J is 2-23; Q¹ are synthons each independently selected from thegroup consisting of (a) phenyl synthons coupled to linkages L at1,4-phenyl positions, (b) aryl synthons other than phenyl synthons, (c)heteroaryl synthons, (d) saturated cyclic hydrocarbon synthons, (e)unsaturated cyclic hydrocarbon synthons, (f) saturated bicyclichydrocarbon synthons, (g) unsaturated bicyclic hydrocarbon synthons, (h)saturated multicyclic hydrocarbon synthons, and (i) unsaturatedmulticyclic hydrocarbon synthons; wherein at least one of Q¹ is a phenylsynthon coupled to linkages L at 1,4-phenyl positions, and wherein ringpositions of each Q¹ which are not coupled to a linkage L aresubstituted with functional groups containing atoms selected from thegroup of C, H, N, O, Si, P, S, B, Al, halogens, and metals from thealkali and alkaline earth groups; Q² is a synthon independently selectedfrom the group consisting of (a) aryl synthons other than phenylsynthons and naphthalene synthons coupled to linkages L at 2,7-naphthylpositions, (b) heteroaryl synthons, (c) saturated cyclic hydrocarbonsynthons other than cyclohexane synthons coupled to linkages L at1,2-cyclohexyl positions, (d) unsaturated cyclic hydrocarbon synthons,(e) saturated bicyclic hydrocarbon synthons, (f) unsaturated bicyclichydrocarbon synthons, (g) saturated multicyclic hydrocarbon synthons,and (h) unsaturated multicyclic hydrocarbon synthons; wherein ringpositions of Q² which are not coupled to an L are substituted withhydrogen or functional groups containing atoms selected from the groupconsisting of C, H, N, O, Si, P, S, B, Al, halogens, and metals from thealkali and alkaline earth groups; L are linkages between the synthonseach independently selected from the group consisting of (a) a condensedlinkage, and (b) a linkage selected from the group consisting of—NRC(O)—, —OC(O)—, —O—, —S—S—, —S—, —NR—, —(CRR′)_(p)—, —CH₂NH—,—C(O)S—, —C(O)O—, —C≡C—, —C≡C—C≡C—, —CH(OH)—, —HC═CH—, —NHC(O)NH—,—NHC(O)O—, —NHCH₂NH—, —NHCH₂CH(OH)CH₂NH—, —N═CH(CH₂)_(p)CH═N—,—CH₂CH(OH)CH₂—, —N═CH(CH₂)_(h)CH═N— where h is 1-4, —CH═N—NH—, —OC(O)O—,—OP(O)(OH)O—, —CH(OH)CH₂NH—, —CH(OH)CH₂—, —CH(OH)C(CH₃)₂C(O)O—,

where G is halogen,

wherein p is 1-6; wherein R and R′ are each independently selected fromthe group of hydrogen and alkyl; wherein linkages L are eachindependently configured in either of two possible configurations,forward and reverse, with respect to the synthons it couples together,if the two configurations are different structures; wherein y is 1 or 2,and Q^(y) are each independently one of the Q¹ or Q² synthons connectedby the linkage.

In other embodiment is a closed ring composition of the formula:

wherein: Q is

J is from 1-22, and n is from 1-24; X and R^(n) are each independentlyselected from the group consisting of functional groups containing atomsselected from the group consisting of C, H, N, O, Si, P, S, B, Al,halogens, and metals from the alkali and alkaline earth groups; Z areeach independently hydrogen or a lipophilic group; L are linkagesbetween synthons each independently selected from the group consistingof (a) a condensed linkage, and (b) a linkage selected from the groupconsisting of —N═CR—, —NRC(O)—, —OC(O)—, —O—, —S—S—, —S—, —NR—,—(CRR′)_(p)—, —CH₂NH—, —C(O)S—, —C(O)O—, —C≡C—, —C≡C—C≡C—, —CH(OH)—,—HC═CH—, —NHC(O)NH—, —NHC(O)O—, —NHCH₂NH—, —NHCH₂CH(OH)CH₂NH—,—N═CHCH₂CH═N— —N═CH(CH₂)_(h)CH═N— where h is 1-4, —CH═N—NH—, —OC(O)O—,—P(O)(OH)₂O—, —CH(OH)CH₂NH—, —CH(OH)CH₂—, —CH(OH)C(CH₃)₂C(O)O—,

wherein p is 1-6; wherein R and R′ are each independently selected fromthe group of hydrogen and alkyl; wherein linkages L are eachindependently configured in either of two possible configurations,forward and reverse, with respect to the synthons it couples together,if the two configurations are different structures.

In one embodiment, X and R^(n) are each independently selected from thegroup consisting of hydrogen, an activated acid, —OH, —C(O)OH, —C(O)H,—C(O)OCH₃, —C(O)Cl, —NRR, —NRRR⁺, —MgX, —Li, —OLi, —OK, —ONa, —SH,—C(O)(CH₂)₂C(O)OCH₃, —NH-alkyl-C(O)CH₂CH(NH₂)CO₂-alkyl, —CH═CH₂,—CH═CHR, —CH═CR₂, 4-vinylaryl, —C(O)CH═CH₂, —NHC(O)CH═CH₂,—C(O)CH═CH(C₆H₅),

—OH, —OC(O)(CH₂)₂C(O)OCH₃, —OC(O)CH═CH₂,

and

—P(O)(OH)(OX), —P(═O)(O⁻)O(CH₂)_(s)NR₃ ⁺;wherein R are each independently selected from the group consisting ofhydrogen and 1-6C alkyl; X is selected from the group consisting of Cl,Br, and I; r is 1-50; and s is 1-4.

In another embodiment is a closed ring composition of the formula:

wherein: Q is

J is from 1-22, and n is from 1-48; X and R^(n) are each independentlyselected from the group consisting of functional groups containing atomsselected from the group consisting of C, H, N, O, Si, P, S, B, Al,halogens, and metals from the alkali and alkaline earth groups; 7 areeach independently hydrogen or a lipophilic group; L are linkagesbetween the synthons each independently selected from the groupconsisting of (a) a condensed linkage, and (b) a linkage selected fromthe group consisting of —NRC(O)—, —OC(O)—, —O—, —S—S—, —S—, —NR—,—(CRR′)_(p)—, —CH₂NH—, —C(O)S—, —C(O)O—, —C≡C—, —C≡C—C≡C—, —CH(OH)—,—HC═CH—, —NHC(O)NH—, —NHC(O)O—, —NHCH₂NH—, —NHCH₂CH(OH)CH₂NH—,—N═CH(CH₂)_(p)CH═N—, —CH₂CH(OH)CH₂—, —N═CH(CH₂)_(h)CH═N— where h is 1-4,—CH═N—NH—, —OC(O)O—, —OP(O)(OH)O—, —CH(OH)CH₂NH—, —CH(OH)CH₂—,—CH(OH)C(CH₃)₂C(O)O—,

where G is halogen,

wherein p is 1-6; wherein R and R′ are each independently selected fromthe group of hydrogen and alkyl; wherein linkages L are eachindependently configured in either of two possible configurations,forward and reverse, with respect to the synthons it couples together,if the two configurations are different structures.

In another embodiment, X and R^(n) are each independently selected fromthe group consisting of hydrogen, an activated acid, —OH, —C(O)OH,—C(O)H, —C(O)OCH₃, —C(O)Cl, —NRR, —NRRR⁺, —MgX, —Li, —OLi, —OK, —ONa,—SH, —C(O)(CH₂)₂C(O)OCH₃, —NH-alkyl-C(O)CH₂CH(NH₂)CO₂-alkyl, —CH═CH₂,—CH═CHR, —CH═CR₂, 4-vinylaryl, —C(O)CH═CH₂, —NHC(O)CH═CH₂,—C(O)CH═CH(C₆H₅),

—OH, —OC(O)(CH₂)₂C(O)OCH₃, —OC(O)CH═CH₂,

and

—P(O)(OH)(OX), —P(═O)(O⁻)O(CH₂)_(s)NR₃ ⁺; wherein R are eachindependently selected from the group consisting of hydrogen and 1-6Calkyl; X is selected from the group consisting of Cl, Br, and I; r is1-50; and s is 1-4.

In another embodiment, the closed ring composition may have the formula:

wherein:

Q is

J is from 1-11, and n is from 1-12; X and R^(n) are each independentlyselected from the group consisting of hydrogen, an activated acid, —OH,—C(O)OH, —C(O)H, —C(O)OCH₃, —C(O)Cl, —NRR, —NRRR⁺, —MgX, —Li, —OLi, —OK,—ONa, —SH, —C(O)(CH₂)₂C(O)OCH₃, —NH-alkyl-C(O)CH₂CH(NH₂)CO₂-alkyl,—CH═CH₂, —CH═CHR, —CH═CR₂, 4-vinylaryl, —C(O)CH═CH₂, —NHC(O)CH═CH₂,—C(O)CH═CH(C₆H₅),

—OH, —OC(O)(CH₂)₂C(O)OCH₃, —OC(O)CH═CH₂,

and

—P(O)(OH)(OX), —P(O)(O⁻)O(CH₂)_(s)NR₃ ⁺;wherein R are each independently selected from the group consisting ofhydrogen and 1-6C alkyl; X is selected from the group consisting of Cl,Br, and I; r is 1-50; and s is 1-4; Z are each independently hydrogen ora lipophilic group; L are linkages between synthons each independentlyselected from the group consisting of (a) a condensed linkage, and (b) alinkage selected from the group consisting of —NRC(O)—, —OC(O)—, —O—,—S—S—, —S—, —NR—, —(CRR′)_(p)—,—CH₂NH—, —C(O)S—, —C(O)O—, —C≡C—, —C≡C—C≡C—, —CH(OH)—, —HC═CH—,—NHC(O)NH—, —NHC(O)O—, —NHCH₂NH—, —NHCH₂CH(OH)CH₂NH—,—N═CH(CH₂)_(p)CH═N—, —CH₂CH(OH)CH₂—, —N═CH(CH₂)_(h)CH═N— where h is 1-4,—CH═N—NH—, —OC(O)O—, —OP(O)(OH)O—, —CH(OH)CH₂NH—, —CH(OH)CH₂—,—CH(OH)C(CH₃)₂C(O)O—,

where G is halogen,

wherein p is 1-6; wherein R and R′ are each independently selected fromthe group of hydrogen and alkyl; wherein linkages L are eachindependently configured in either of two possible configurations,forward and reverse, with respect to the synthons it couples together,if the two configurations are different structures.

In another embodiment the closed ring composition may have the formula:

wherein:Q is

J is from 1-11, and n is from 1-12;X and R^(n) are each independently selected from the group consisting ofhydrogen, an activated acid, —OH, —C(O)OH, —C(O)H, —C(O)OCH₃, —C(O)Cl,—NRR, —NRRR⁺, —MgX, —Li, —OLi, —OK, —ONa, —SH, —C(O)(CH₂)₂C(O)OCH₃,—NH-alkyl-C(O)CH₂CH(NH₂)CO₂-alkyl, —CH═CH₂, —CH═CHR, —CH═CR₂,4-vinylaryl, —C(O)CH═CH₂, —NHC(O)CH═CH₂, —C(O)CH═CH(C₆H₅),

—OH, —OC(O)(CH₂)₂C(O)OCH₃, —OC(O)CH═CH₂,

and

—P(O)(OH)(OX), —P(═O)(O⁻)O(CH₂)_(s)NR₃ ⁺; wherein R are eachindependently selected from the group consisting of hydrogen and 1-6Calkyl; X is selected from the group consisting of Cl, Br, and I; r is1-50; and s is 1-4; Z are each independently hydrogen or a lipophilicgroup; L are linkages between the synthons each independently selectedfrom the group consisting of (a) a condensed linkage, and (b) a linkageselected from the group consisting of —NRC(O)—, —OC(O)—, —O—, —S—S—,—S—, —NR—, —(CRR′)_(p)—, —CH₂NH—, —C(O)S—, —C(O)O—, —C≡C—, —C≡C—C≡C—,—CH(OH)—, —HC═CH—, —NHC(O)NH—, —NHC(O)O—, —NHCH₂NH—, —NHCH₂CH(OH)CH₂NH—,—N═CH(CH₂)_(p)CH═N—, —CH₂CH(OH)CH₂—, —N═CH(CH₂)_(h)CH═N— where h is 1-4,—CH═N—NH—, —OC(O)O—, —OP(O)(OH)O—, —CH(OH)CH₂NH—, —CH(OH)CH₂—,—CH(OH)C(CH₃)₂C(O)O—,

where G is halogen,

wherein p is 1-6; wherein R and R′ are each independently selected fromthe group of hydrogen and alkyl; wherein linkages L are eachindependently configured in either of two possible configurations,forward and reverse, with respect to the synthons it couples together,if the two configurations are different structures.

In another embodiment, the closed ring composition may have the formula:

wherein:Q is

J is from 1-11, and n is from 1-12; X is —NX¹— or —CX²X³, where X¹ isselected from the group consisting of an amino acid residue,—CH₂C(O)CH₂CH(NH₂)CO₂-alkyl, and —C(O)CH═CH₂; X² and X³ are eachindependently selected from the group consisting of hydrogen, —OH, —NH₂,—SH, —(CH₂)_(t)OH, —(CH₂)_(t)NH₂ and —(CH₂)_(t)SH, wherein t is 1-4, andX² and X³ are not both hydrogen; R^(n) are each independently selectedfrom the group consisting of hydrogen, an activated acid, —OH, —C(O)OH,—C(O)H, —C(O)OCH₃, —C(O)Cl, —NRR, —NRRR⁺, —MgX, —Li, —OLi, —OK, —ONa,—SH, —C(O)(CH₂)₂C(O)OCH₃, —NH-alkyl-C(O)CH₂CH(NH₂)CO₂-alkyl, —CH═CH₂,—CH═CHR, —CH═CR₂, 4-vinylaryl, —C(O)CH═CH₂, —NHC(O)CH═CH₂,—C(O)CH═CH(C₆H₅),

—OH, —OC(O)(CH₂)₂C(O)OCH₃, —OC(O)CH═CH₂,

and

—P(O)(OH)(OX), —P(═O)(O⁻)O(CH₂)_(s)NR₃ ⁺;wherein R are each independently selected from the group consisting ofhydrogen and 1-6C alkyl; X is selected from the group consisting of Cl,Br, and I; r is 1-50; and s is 1-4; Z are each independently hydrogen ora lipophilic group; L are linkages between synthons each independentlyselected from the group consisting of (a) a condensed linkage, and (b) alinkage selected from the group consisting of —NRC(O)—, —OC(O)—, —O—,—S—S—, —S—, —NR—, —(CRR′)_(p)—, —CH₂NH—, —C(O)S—, —C(O)O—, —C≡C—,—C≡C—C≡C—, —CH(OH)—, —HC═CH—, —NHC(O)NH—, —NHC(O)O—, —NHCH₂NH—,—NHCH₂CH(OH)CH₂NH—, —N═CH(CH₂)_(p)CH═N—, —CH₂CH(OH)CH₂—,—N═CH(CH₂)_(h)CH═N— where h is 1-4, —CH═N—NH—, —OC(O)O—, —OP(O)(OH)O—,—CH(OH)CH₂NH—, —CH(OH)CH₂—, —CH(OH)C(CH₃)₂C(O)O—,

where G is halogen,

wherein p is 1-6; wherein R and R′ are each independently selected fromthe group of hydrogen and alkyl; wherein linkages L are eachindependently configured in either of two possible configurations,forward and reverse, with respect to the synthons it couples together,if the two configurations are different structures.

In another embodiment the closed ring structure may have the formula:

wherein:Q is

J is from 1-11, and n is from 1-12; X and R^(n) are each independentlyselected from the group consisting of hydrogen, an activated acid, —OH,—C(O)OH, —C(O)H, —C(O)OCH₃, —C(O)Cl, —NRR, —NRRR⁺, —MgX, —Li, —OLi, —OK,—ONa, —SH, —C(O)(CH₂)₂C(O)OCH₃, —NH-alkyl-C(O)CH₂CH(NH₂)CO₂-alkyl,—CH═CH₂, —CH═CHR, —CH═CR₂, 4-vinylaryl, —C(O)CH═CH₂, —NHC(O)CH═CH₂,—C(O)CH═CH(C₆H₅),

—OH, —OC(O)(CH₂)₂C(O)OCH₃, —OC(O)CH═CH₂,

and

—P(O)(OH)(OX), —P(═O)(O⁻)O(CH₂)_(s)NR₃ ⁺; wherein R are eachindependently selected from the group consisting of hydrogen and 1-6Calkyl; X is selected from the group consisting of Cl, Br, and I; r is1-50; and s is 1-4; Z and Y are each independently hydrogen or alipophilic group; L are linkages between the synthons each independentlyselected from the group consisting of (a) a condensed linkage, and (b) alinkage selected from the group consisting of —NRC(O)—, —OC(O)—, —O—,—S—S—, —S—, —NR—, —(CRR′)_(p)—, —CH₂NH—, —C(O)S—, —C(O)O—, —C≡C—,—C≡C—C≡C—, —CH(OH)—, —HC═CH—, —NHC(O)NH—, —NHC(O)O—, —NHCH₂NH—,—NHCH₂CH(OH)CH₂NH—, —N═CH(CH₂)_(p)CH═N—, —CH₂CH(OH)CH₂—,—N═CH(CH₂)_(h)CH═N— where h is 1-4, —CH═N—NH—, —OC(O)O—, —OP(O)(OH)O—,—CH(OH)CH₂NH—, —CH(OH)CH₂—, —CH(OH)C(CH₃)₂C(O)O—,

where G is halogen,

wherein p is 1-6;wherein R and R′ are each independently selected from the group ofhydrogen and alkyl; wherein linkages L are each independently configuredin either of two possible configurations, forward and reverse, withrespect to the synthons it couples together, if the two configurationsare different structures.

In another embodiment, the closed ring structure may have the formula:

wherein:Q is

J is from 1-11, and n is from 1-12; X and R^(n) are each independentlyselected from the group consisting of hydrogen, an activated acid, —OH,—C(O)OH, —C(O)H, —C(O)OCH₃, —C(O)Cl, —NRR, —NRRR⁺, —MgX, —Li, —OLi, —OK,—ONa, —SH, —C(O)(CH₂)₂C(O)OCH₃, —NH-alkyl-C(O)CH₂CH(NH₂)CO₂-alkyl,—CH═CH₂, —CH═CHR, —CH═CR₂, 4-vinylaryl, —C(O)CH═CH₂, —NHC(O)CH═CH₂,—C(O)CH═CH(C₆H₅),

—OH, —OC(O)(CH₂)₂C(O)OCH₃, —OC(O)CH═CH₂,

and

—P(O)(OH)(OX), —P(═O)(O⁻)O(CH₂)_(s)NR₃ ⁺;wherein R are each independently selected from the group consisting ofhydrogen and 1-6C alkyl; X is selected from the group consisting of Cl,Br, and I; r is 1-50; and s is 1-4; Z and Y are each independentlyhydrogen or a lipophilic group; L are linkages between synthons eachindependently selected from the group consisting of (a) a condensedlinkage, and (b) a linkage selected from the group consisting of—NRC(O)—, —OC(O)—, —O—, —S—S—, —S—, —NR—, —(CRR′)_(p)—, —CH₂NH—,—C(O)S—, —C(O)O—, —C≡C—, —C≡C—C≡C—, —CH(OH)—, —HC═CH—, —NHC(O)NH—,—NHC(O)O—, —NHCH₂NH—, —NHCH₂CH(OH)CH₂NH—, —N═CH(CH₂)_(p)CH═N—,—CH₂CH(OH)CH₂—, —N═CH(CH₂)_(h)CH═N— where h is 1-4, —CH═N—NH—, —OC(O)O—,—OP(O)(OH)O—, —CH(OH)CH₂NH—, —CH(OH)CH₂—, —CH(OH)C(CH₃)₂C(O)O—,

where G is halogen,

and

wherein p is 1-6; wherein R and R′ are each independently selected fromthe group of hydrogen and alkyl; wherein linkages L are eachindependently configured in either of two possible configurations,forward and reverse, with respect to the synthons it couples together,if the two configurations are different structures.

In one variation, a macrocyclic module may be a closed ring compositionof the formula:

wherein: the closed ring comprises a total of from three to twenty-foursynthons Q; J is 2-23; Q¹ are synthons each independently selected fromthe group consisting of (a) aryl synthons, (b) heteroaryl synthons, (c)saturated cyclic hydrocarbon synthons, (d) unsaturated cyclichydrocarbon synthons, (e) saturated bicyclic hydrocarbon synthons, (f)unsaturated bicyclic hydrocarbon synthons, (g) saturated multicyclichydrocarbon synthons, and (h) unsaturated multicyclic hydrocarbonsynthons; wherein ring positions of each Q¹ which are not coupled to alinkage L are substituted with hydrogen or functional groups containingatoms selected from the group of C, H, N, O, Si, P, S, B, Al, halogens,and metals from the alkali and alkaline earth groups; Q² is a synthonindependently selected from the group consisting of (a) aryl synthons,(b) heteroaryl synthons, (c) saturated cyclic hydrocarbon synthons, (d)unsaturated cyclic hydrocarbon synthons, (e) saturated bicyclichydrocarbon synthons, (f) unsaturated bicyclic hydrocarbon synthons, (g)saturated multicyclic hydrocarbon synthons, and (h) unsaturatedmulticyclic hydrocarbon synthons; wherein ring positions of Q² which arenot coupled to an L are substituted with hydrogen or functional groupscontaining atoms selected from the group consisting of C, H, N, O, Si,P, S, B, Al, halogens, and metals from the alkali and alkaline earthgroups; L are linkages between the synthons each independently selectedfrom the group consisting of synthon-synthon, —NRC(O)—, —OC(O)—, —O—,—S—S—, —S—, —NR—, —(CRR′)_(p)—, —CH₂NH—, —C(O)S—, —C(O)O—, —C≡C—,—C≡C—C≡C—, —CH(OH)—, —HC═CH—, —NHC(O)NH—, —NHC(O)O—, —NHCH₂NH—,—NHCH₂CH(OH)CH₂NH—, —N═CH(CH₂)_(p)CH═N—, —CH₂CH(OH)CH₂—,—N═CH(CH₂)_(h)CH═N— where h is 1-4, —CH═N—NH—, —OC(O)O—, —OP(O)(OH)O—,—CH(OH)CH₂NH—, —CH(OH)CH₂—, —CH(OH)C(CH₃)₂C(O)O—,

wherein p is 1-6; wherein R and R′ are each independently selected fromthe group of hydrogen and alkyl; wherein the linkages L are eachindependently configured with respect to the Q¹ and Q² synthons, each Lhaving either of its two possible configurations with respect to thesynthons it couples together, the forward and reverse configurations ofthe linkage with respect to the immediately adjacent synthons to whichit couples, for example, Q¹ _(a)-NHC(O)-Q¹ _(b) and Q¹ _(a)-C(O)NH-Q¹_(b), if the two configurations are isomerically different structures.Synthons Q¹, when independently selected, may be any cyclic synthon asdescribed, so that the J synthons Q¹ may be found in the closed ring inany order, for example,cyclohexyl-1,2-phenyl-piperidinyl-1,2-phenyl-1,2-phenyl-cyclohexyl, andso on, and the J linkages L may also be independently selected andconfigured in the closed ring. The macrocyclic modules represented andencompassed by the formula include all stereoisomers of the synthonsinvolved, so that a wide variety of stereoisomers of the macrocyclicmodule are included for each closed ring composition of synthons.

Methods for making a macrocyclic module composition may comprise, forexample: (a) providing a plurality of a first cyclic synthon; (b)contacting a plurality of a second cyclic synthon with the first cyclicsynthons; (c) isolating the macrocyclic module composition. In someembodiments, the method may further comprise contacting a linkermolecule with the mixture in (a) or (b).

Another method of preparing a composition for transporting a selectedspecies through the composition comprises: selecting a first cyclicsynthon, wherein the first cyclic synthon is substituted with at leastone functional group comprising an functional group containing atomsselected from the group consisting of C, H, N, O, Si, P, S, B, Al,halogens, and metals from the alkali and alkaline earth groups;selecting from two to about twenty-three additional cyclic synthons;incorporating the first cyclic synthon and the additional cyclicsynthons into a macrocyclic module composition comprising: from three toabout twenty-four cyclic synthons coupled to form a closed ring defininga pore; wherein the at least one functional group of the first cyclicsynthon is located at the pore of the macrocyclic module composition andis selected to transport the selected species through the pore.

Another method for making a macrocyclic module composition comprises:(a) providing a plurality of a first cyclic synthon; (b) contacting aplurality of a second cyclic synthon with the first cyclic synthons; (c)contacting a plurality of the first cyclic synthon with the mixture from(b).

Another method for making a macrocyclic module composition comprises:(a) providing a plurality of a first cyclic synthon; (b) contacting aplurality of a second cyclic synthon with the first cyclic synthons; (c)contacting a plurality of a third cyclic synthon with the mixture from(b).

In some embodiments, the method may further comprise supporting a cyclicsynthon or coupled synthons on a solid phase.

Another method for making a macrocyclic module composition comprises:(a) contacting a plurality of cyclic synthons with a metal complextemplate; (b) isolating the macrocyclic module composition.

A macrocyclic module may include functional groups for coupling themacrocyclic module to a solid surface, substrate, or support. Examplesof functional groups of macrocyclic modules which can be used to coupleto a substrate or surface include amine, carboxylic acid, carboxylicester, benzophenone and other light activated crosslinkers, alcohol,glycol, vinyl, styryl, olefin styryl, epoxide, thiol, magnesium halo orGrignard, acrylate, acrylamide, diene, aldehyde, and mixtures thereof.These functional groups may be coupled to the closed ring of themacrocyclic module, and may optionally be attached by a spacer group.Examples of solid surfaces include metal surfaces, ceramic surfaces,polymer surfaces, semiconductor surfaces, silicon wafer surfaces,alumina surfaces, and so on. Examples of functional groups ofmacrocyclic modules which can be used to couple to a substrate orsurface further include those described in the left hand column ofTables 5 and 6. Methods of initiating coupling of the modules to thesubstrate include chemical, thermal, photochemical, electrochemical, andirradiative methods.

Examples of spacer groups include polyethylene oxides, polypropyleneoxides, polysaccharides, polylysines, polypeptides, poly(amino acids),polyvinylpyrrolidones, polyesters, polyvinylchlorides, polyvinylidenefluorides, polyvinylalcohols, polyurethanes, polyamides, polyimides,polysulfones, polyethersulfones, polysulfonamides, and polysulfoxides.

Macrocyclic Module Pores

An individual macrocyclic module may include a pore in its structure.The size of the pore may determine the size of molecules or otherspecies which can pass through the macrocyclic module. The size of apore in a macrocyclic module may depend on the structure of the synthonsused to make the macrocyclic module, the linkages between synthons, thenumber of synthons in a module, the structure of any linker moleculesused to make the macrocyclic module, and other structural features ofthe macrocyclic module whether inherent in the preparation of themacrocyclic module or added in later steps or modifications.Stereoisomerism of macrocyclic modules may also be used to regulate thesize of a pore of a macrocyclic module by variation of the stereoisomerof each synthon used to prepare the closed ring of the macrocyclicmodule.

The dimension of a pore in a macrocyclic module may be varied bychanging the combination of synthons used to form the macrocyclicmodule, or by varying the number of synthons in the closed ring. Thedimension of a pore may also be varied by substituents on the synthonsor linkages. The pore may therefore be made large enough or small enoughto achieve an effect on transport of species through the pore. Specieswhich may be transported through the pore of a macrocyclic moduleinclude atoms, molecules, biomolecules, ions, charged particles, andphotons.

The size of a species may not be the sole determinant of whether it willbe able to pass through a pore of a macrocyclic module. Groups ormoieties located in or near the pore structure of a macrocyclic modulemay regulate or affect transport of a species through the pore byvarious mechanisms. For example, transport of a species through the poremay be affected by groups of the macrocyclic module which interact withthe species, by ionic or other interaction, such as chelating groups, orby complexing the species. For example, a charged group such as acarboxylate anion or ammonium group may couple an oppositely-chargedspecies and affect its transport. Substituents of synthons in amacrocyclic module may affect the passage of a species through the poreof the macrocyclic module. Groups of atoms which render the pore of amacrocyclic module more or less hydrophilic or lipophilic may affecttransport of a species through the pore. An atom or group of atoms maybe located within or proximate to a pore to sterically slow or block thepassage of a species through the pore. For example, hydroxyl or alkoxygroups may be coupled to a cyclic synthon and located in the pore of thestructure of the macrocyclic module, or may be coupled to a linkagebetween synthons and located in the pore. A wide range of functionalgroups may be used to sterically slow or block the passage of a speciesthrough the pore, including functional groups containing atoms selectedfrom the group consisting of C, H, N, O, Si, P, S, B, Al, halogens, andmetals from the alkali and alkaline earth groups. Blocking and slowingpassage of a species through the pore may involve reducing the dimensionof the pore by steric blocking, as well as slowing the passage ofspecies by creating a path through the pore which is not linear, andproviding interaction between the functional group and the species toslow transport. The stereochemical structure of the portion of themacrocyclic module which defines the pore and its interior may alsoaffect transport. Any groups or moieties which affect transport of aspecies through the pore of a macrocyclic module may be introduced aspart of the synthons used to prepare the macrocyclic module, or may beadded later by various means. For example, S7-1 could be reacted withClC(O)(CH₂)₂C(O)OCH₂CH₃ to convert the phenol groups to succinyl estergroups. Further, molecular dynamical motion of the synthons and linkagesof a partly flexible macrocyclic module may affect transport of aspecies through the pore of the module. Transport behavior may not bedescribed solely by the structure of the macrocyclic module itself sincethe presence of the species which is to be transported through the poreaffects the flexibility, conformation, and dynamical motions of amacrocyclic module. In general, solvent may also affect transport ofsolutes through a pore.

Macrocyclic modules and arrays of macrocyclic modules may be useful insize exclusion separations, ion separation, gas separation, separationof enantiomers, small molecule separation, water purification,filtration of bacteria, fungi, or viruses, sewage treatment, and toxinremoval, among other uses.

The following examples further describe and demonstrate variationswithin the scope of the present invention. All examples described inthis specification, both in the description above and the examplesbelow, are given solely for the purpose of illustration and are not tobe construed as limiting the present invention. While there have beendescribed illustrative variations of this invention, those skilled inthe art will recognize that they may be changed or modified withoutdeparting from the spirit and scope of this invention, and it isintended to cover all such changes, modifications, and equivalentarrangements that fall within the true scope of the invention as setforth in the appended claims.

All documents referenced herein, including applications for patent,patent references, publications, articles, books, and treatises, arespecifically incorporated by reference herein in their entirety.

EXAMPLES

Reagents were obtained from Aldrich Chemical Company and VWR ScientificProducts. Reactions were carried out under nitrogen or argon atmosphereunless otherwise noted. Solvent extracts of aqueous solutions were driedover anhydrous Na₂SO₄. Solutions were concentrated under reducedpressure using a rotary evaporator.

Example 1 Derivatization of Silicon Substrates with(3-aminopropyl)triethoxysilane (APTES)

SiO₂ substrates were first sonicated in a piranha solution (3:1 ratio ofH₂SO₄:30% H₂O₂) for 15 minutes. This was followed by a 15 min sonicationin Milli-Q water (>18 MΩ-cm). The derivatization step was done in aglove bag under a N₂ atmosphere. 0.05 mL APTES and 0.05 mL pyridine wereadded to 9 mL of toluene. Immediately following mixing, the freshlycleaned SiO₂ substrates were immersed in the APTES solution for 10 min.Substrates were washed with copious amounts of toluene and then driedwith N₂. Deposited APTES films showed a range of thickness values from0.8 to 1.3 nm.

Example 2

Amphiphilic modules Hexamer 1a were dissolved in HPLC-grade chloroformat a concentration of approximately 1 mg/ml. The chloroform solution wasapplied to a water (Millipore Milli-Q) surface in an L-B trough (KSV,Helsinki). The chloroform was allowed to evaporate, leaving theamphiphilic modules on the surface of the water with their hydrophilicgroups immersed in the water and their lipophilic groups in the air. Thetemperature in the system was controlled (approximately ±0.2° C.). Thebarriers of the L-B trough were slowly compressed (1-10 mm/min). Thesurface pressure was monitored using the Wilhelmy procedure during filmcompression. The shape of the isotherm confirmed that the module formeda Langmuir film on the surface of the water.

The scheme for this preparation is illustrated in FIGS. 1A and 1B. 25 μlof a 1 mg/ml solution of Hexamer 1 dh in chloroform were spread on a 2mg/ml diethyl malonimidate (DEM) in water subphase (pH 8.8, T=22° C.).After waiting for 19 minutes to allow for spreading solvent evaporation,the monolayer was compressed and held at 5 mN/m. The subphase was thenheated to 40° C. and held for approximately 60 minutes. A rigidifiedsolid nanofilm was formed on the surface of the subphase, which appeareduniform and homogeneous in a Brewster Angle microscope image. Whentouched with a probe, the film was cracked, as shown by the BrewsterAngle microscope image obtained for the film directly on the surface ofthe subphase, illustrated in FIG. 6A. This indicates that the nanofilmwas highly cross-linked. Mass spectral analysis of the rigid nanofilmcould not be performed because the solid nanofilm clogged the input ofthe instrument.

Example 3 Hexamer 1 dh with DEM (Langmuir-Blodgett Deposition)

The scheme for this preparation is illustrated in FIGS. 1A and 1B. 25 μlof a 1 mg/ml solution of Hexamer 1 dh in chloroform were spread on a 2mg/ml diethyl malonimidate (DEM) in water subphase (pH 8.8, T=22° C.).After waiting for 19 minutes to allow for spreading solvent evaporation,the monolayer was compressed and held at 5 mN/m. The subphase was thenheated to 40° C. and held for approximately 60minutes. A rigidifiedsolid nanofilm was formed on the surface of the subphase, which appeareduniform and homogeneous in a Brewster Angle microscope image. Whentouched with a probe, the film was cracked, as shown by the BrewsterAngle microscope image obtained for the film directly on the surface ofthe subphase, illustrated in FIG. 6A. This indicates that the nanofilmwas highly cross-linked. Mass spectral analysis of the rigid nanofilmcould not be performed because the solid nanofilm clogged the input ofthe instrument.

Another monolayer prepared as above was transferred to a Si wafer (madehydrophilic by treating with piranha solution, 3:1 ratio of H₂SO₄/30%H₂O₂, for 10 min) by Langmuir-Blodgett deposition. The substrate wastranslated through the air-water interface at a speed of 0.3 mm/min.Imaging ellipsometry, illustrated in FIG. 6B, revealed shattered solidislands of film on the silicon substrate with heights of approximately35 Å, which includes the nanofilm and an APTES coating of the substrate.

The isobaric creep of the solid nanofilm is illustrated in FIG. 7. ALangmuir film which has individual molecules oriented on a surface in acondensed state, but in which there is no coupling between the moleculeswill spread out or decompress when the force of compression is released.By comparison, the solid nanofilm of coupled modules retained its shapeand film strength over time, even in the presence of solvent, as shownin FIG. 7. The upper line shows the isobaric creep of the nanofilmprepared in the presence of DEM cross-linker, and the lower line showsthe isobaric creep of the nanofilm prepared without crosslinker.

Example 4 Hexamer 1 dh with DEM (Langmuir-Blodgett Deposition)

The presence of interlinking of modules was detected by FourierTransform Infrared Spectroscopy (FTIR). The FTIR spectra of the nanofilmof Example 3 are illustrated in FIG. 8. In FIG. 8A, the FTIR spectra ofthe nanofilm and the Hexamer 1 dh are illustrated. In FIG. 8B, the FTIRspectra of DEM and DEM in the subphase after 24 hours are illustrated.Changes in the FTIR show that there was coupling between Hexamer 1 dhand DEM.

Example 5 Hexamer 1 dh with DEM (Langmuir-Schaefer Deposition)

25 μl of a 1 mg/ml solution of Hexamer 1 dh in chloroform were spread ona 2 mg/ml diethyl malonimidate subphase (pH 8.8, T=22° C.). Afterwaiting 15 minutes to allow for spreading solvent evaporation, themonolayer was compressed and held at 5 mN/m. The monolayer wastransferred to a Si wafer (made hydrophilic by treating with piranhasolution for 10 minutes) by a Langmuir-Schaefer deposition. Imagingellipsometry revealed an intact film on the silicon substrate with aheight of approximately 21 Å, as illustrated in FIG. 9, in which asingle fracture of the nanofilm is illustrated.

Example 6 Mannich Reaction with Octadecylamine

35 μl of a 1 mg/ml solution of octadecylamine in chloroform were spreadon a 1% formaldehyde subphase (pH 3, T=22° C.). After waiting 15 minutesto allow for spreading solvent evaporation, the monolayer was compressedand held at 20 mN/m. At this surface pressure, the film exhibited asteady loss in area for approximately 80minutes, after which the filmarea increased. After a total of 130 minutes at the air-water interface,the film was extracted by manual Langmuir-Blodgett depositions. Briefly,after dipping a Si wafer through the air-water interface, the wafer wasshaken in chloroform to remove the deposited material, and the processwas repeated. Mass spectrometry (ESI mode) was then performed on thechloroform solution. The structures of the linkages formed in thenanofilm upon coupling the amphiphiles as detected in the mass spectrumare illustrated FIGS. 4B, 4D and 4F.

Example 7 Surface Attachment with Reactive Ester Groups

First, the APTES modified silicon substrates were lowered into a pH 7,22° C. aqueous subphase. 160 mL of methylheptadecanoate (MHD) (1 mg/mLCHCl₃ solution) was spread at the air/water interface. After 10 min thefilm was compressed to 38 mN/m at a rate of 3 mm/min. Upon reaching 38mN/m the substrates were raised out of the subphase (while maintainingthe surface pressure at 38 mN/m) at a rate of 1 mm/min resulting indeposition of one layer of MHD. Following deposition, some samples wereheated at 70° C. for 3.5 hr to induce reaction between the surface aminegroups (APTES) and the ester groups of MHD to form amide linkages.Samples were sonicated in CHCl₃ following the thermal cure to determinethe extent of surface attachment. If the film did not react with thesurface this treatment should result in the removal of the film.

Ellipsometric images of the substrate are shown in FIG. 10. In FIG. 10Ais shown the substrate after film deposition. In FIG. 10B is shown thesubstrate after film deposition and heating at 70° C. FIG. 10B indicatesthat some dewetting occurred during heating, and that nanofilm remainedon the substrate. In FIG. 10C is shown the substrate after filmdeposition, after heating at 70° C., and after treatment with CHCl₃,again indicating that nanofilm remained on the substrate.

Example 8 Surface Attachment with Reactive Acryl Groups

First, SiO₂ substrates were derivatized with a layer ofmethylacryloxymethyltrimethoxysilane (MAOMTMOS) using the same procedureas described in the derivatization with APTES (Example 6). Thesubstrates were then lowered into a pH 5, 22° C. aqueous subphase. 170mL of N-octadecylacrylamide (ODAA) (1 mg/mL CHCl₃ solution) was spreadat the air/water interface. After 10 min the film was compressed to 35mN/m at a rate of 2 mm/min. Upon reaching 35 mN/m the substrates wereraised out of the subphase at a rate of 2 mm/min resulting in thedeposition of one layer of ODAA. Following deposition some samples wereirradiated (254 nm) for 40 or 220 min. to induce coupling between thesurface acryl groups (MAOMTMOS) and the acryl groups of ODAA. Sampleswere sonicated in CHCl₃ following the UV cure to determine the extent ofsurface attachment. If the film did not react with the surface thistreatment would have resulted in removal of the film.

Ellipsometric images of the substrate are shown in FIG. 11. In FIG. 11Ais shown the substrate after film deposition and exposure to UVirradiation at 254 nm for 40 minutes. In FIG. 11B is shown the substrateafter film deposition, irradiation, and after treatment with CHCl₃. FIG.11B shows that there was monolayer coupling to the substrate.

Example 9

A unique structure of the nanofilm of Hexamer 1 dh of Example 3 isillustrated in FIG. 12, in which interlinking is by amide linkages ofthe modules through the cyclohexyl synthons. The approximate dimensionsof the modular and interstitial pores of the structure are 14 Å², 25 Å²and 40 Å². The fully minimized structure was obtained by MM+ molecularmechanics.

Example 10

Octamer 5jh-aspartic is formed in a condensed Langmuir film and heatedto a temperature sufficient to initiate coupling of the modules throughamide linkages to form a nanofilm. A unique structure of a nanofilm ofoctamer 5jh-aspartic is illustrated in FIG. 13, in which interlinking isby aspartic amide linkages of the modules through the piperidinesynthons. The approximate dimensions of the pores of the structure arefrom 119 Å² to 200 Å². The fully minimized structure was obtained by MM+molecular mechanics.

Example 11

First, two SiO₂ substrate were derivatized with a layer ofacryloxy-propyltrimethoxysilane (AOPTMOS) using the same procedure asdescribed in the derivatization with APTES (see example 1). Thesesubstrates, as well as an unmodified SiO₂ substrate, were then loweredinto a H₂O subphase that was maintained at 22° C. Subsequently, theHexamer 1jh-AC (1 mg/mL CHCl₃ solution) was spread at the air/waterinterface. After 10 min the film was compressed to 30 mN/m at a rate of4 mm/min. Upon reaching a surface pressure of 30 mN/m, the Langmuir filmwas irradiated with 254 nm light from a distance of 1.5 inches (1350μW/cm² at 3 inches) for 30 min. Subsequently, the substrates were raisedout of the subphase at a rate of 1 mm/min resulting in the deposition ofone layer of cross-linked Hexamer 1jh-AC. Following deposition, thesample deposited on the AOPTMOS substrate was irradiated (254 nm) for 30min to induce coupling between the surface acryl groups (AOPTMOS) andthe acryl groups of Hexamer 1jh-AC. All samples were then examined byellipsometry to determine film thickness values. Finally, all sampleswere sonicated in CHCl₃ to determine the extent of surface attachment.If the film did not react with the surface this treatment should resultin removal of the film. The corresponding Langmuir trough area vs. time(during irradiation) graph and ellipsometric images of the depositedfilms are shown in FIGS. 14 and 15, respectively. The ellipsometricimages for the film deposited on the MAOMTMOS modified substrate clearlyshows that the films are still present after CHCl₃ sonication, andtherefore indicate that surface attachment occurred. Conversely, when UVlight was not used after deposition on the MAOMTMOS modified substrateor when the film was deposited on silicon, the ellipsometric imagesindicate that surface attachment did not occur.

Additionally, FTIR data reveal the loss of vinyl bands upon UV exposureof the Hexamer 1jh-AC (FIG. 16), indicating crosslinking between Hexamer1jh-AC and the AOPTMOS-derivatized substrate.

Example 12

The filtration function of a membrane may be described in terms of itssolute rejection profile. The filtration function of some nanofilmmembranes is exemplified in Tables 7-8.

TABLE 7 Example filtration function of a G-membrane MOLECULAR SOLUTEWEIGHT PASS/NO PASS Albumin 68 kDa NP Ovalbumin 44 kDa P Myoglobin 17kDa P β₂-Microglobulin 12 kDa P Insulin 5.2 kDa P Vitamin B₁₂ 1350 Da PUrea, H₂O, ions <1000 Da P

TABLE 8 Example filtration function of a T-membrane MOLECULAR SOLUTEWEIGHT PASS/NO PASS β₂-Microglobulin 12 kDa NP Insulin 5.2 kDa NPVitamin B₁₂ 1350 Da NP Glucose 180 Da NP Creatinine 131 Da NP H₂PO₄ ⁻,HPO₄ ²⁻ ≈97 Da NP HCO₃ ⁻ 61 Da NP Urea 60 Da NP K+ 39 Da P Na+ 23 Da P

The passage or exclusion of a solute is measured by its clearance, whichreflects the portion of solute that actually passes through themembrane. The no pass symbol in Tables 7-8 indicates that the solute ispartly excluded by the nanofilm, sometimes less than 90% rejection,often at least 90% rejection, sometimes at least 98% rejection. The passsymbol indicates that the solute is partly cleared by the nanofilm,sometimes less than 90% clearance, often at least 90% clearance,sometimes at least 98% clearance.

A membrane is impermeable to a species if it has a very low clearance(for example, less than about 5%, less than about 3%) for the species,or if it has very high rejection for the species (for example, greaterthan about 95%, greater than about 98%).

Example 13

The dimensions of module pores may be measured by electrical conductancein a voltage-clamped lipid bilayer test. Modules are dissolved into aphosphatidylcholine-phosphatidylethanolamine lipid bilayer. On one sideof the bilayer is placed a solution containing a test cationic species.On the other side is placed a solution containing a cationic speciesknown to be able to pass through the module pore. Anions required forcharge neutrality are selected such that they will not pass through themodule pore. When a positive potential is created in the solution on theside of the lipid bilayer containing the test species, if the testcations are of such a size that they cannot pass through the pores inthe modules, no current will be detected. The voltage is then reversedto create a positive potential on the side of the lipid bilayer havingthe solution containing the cationic species known to be able totraverse the pore. Observation of the expected current confirms theintegrity of the lipid bilayer and the availability of the module poresas transporters of cations of the known size and smaller.

The selective permeability of macrocyclic modules was tested using thevoltage-clamped bilayer method, as shown in Table 9. The “+” symbolindicates permeation of the solute, the “−” symbol indicates rejectionof the solute. Permeation and rejection are indicators of clearance. Theclamp voltage was 50 mV.

TABLE 9 Examples of permeation of macrocyclic modules Radius Solute(VdW, Å) Hexamer 1a Hexamer 1jh Li⁺ 0.8 − + Na⁺ 1.0 + + K⁺ 1.3 + + Cs⁺1.7 + + Ca² ⁺ 1.0 + + Mg² ⁺ 0.7 − + NH₄ ⁺ 1.9 + + MeNH₃ ⁺ 2.0 + + EtNH₃⁺ 2.6 − + NMe₄ ⁺ 2.6 − + Aminoguanidinium 3.1 − + NEt₄ ⁺ 3.9 − + Choline3.8 − + Glucosamine 4.2 − + N(n-Pr)₄ ⁺ − + N(n-Bu)₄+ − +

Example 14

Selective filtration and relative clearance of solutes is exemplified inTable 10. In Table 10, the heading “high permeability” indicates aclearance of greater than about 70-90% of the solute. The heading“medium permeability” indicates a clearance of less than about 50-70% ofthe solute. The heading “low permeability” indicates a clearance of lessthan about 10-30% of the solute.

TABLE 10 Clearance of solutes by nanofilms Nanofilm high permeabilitymedium permeability low permeability Hexamer 1a H₂O, Na⁺, K⁺, Cs⁺ Ca²⁺,Mg²⁺, phosphate Glucose, Li⁻, urea, creatinine water H₂O Glucose, Na⁺,K⁺, Ca²⁺, Mg²⁻, Li⁻, urea, nanofilm phosphate creatinine ion H₂O, Na⁺,K⁺, phosphate Glucose Ca²⁺, Mg²⁺, Li⁻, urea, nanofilm creatinine glucoseH₂O, Na⁺, K⁺, Glucose Phosphate Ca²⁺, Mg²⁻, Li⁺, urea, nanofilmcreatinine G H₂O, Na⁺, K⁺, phosphate, Vitamin B₁₂, Insulin, β₂ Myglobin,Ovalbumin, nanofilm Glucose, Ca²⁺, Mg²⁺, Li⁺, Microglobulin Albumin,urea, creatinine gas He, H₂ — H₂O and larger, liquids in nanofilmgeneral anion Cl⁻ HCO₃ ⁻, Phosphate — nanofilm

Example 15

The approximate diameter of various species to be considered in afiltration process are illustrated in Table 11:

solute molecular weight (Da) diameter (Å) virus 10⁶ 133 immunoglobulin G(IgG) 10⁵ 60 albumin 50 × 10⁴ 50 β₂-Microglobulin 10³ 13 urea 60 — Na⁺23 —Synthon and Macrocyclic Module Synthesis Methods

All chemical structures illustrated and described in this specification,both in the description above and the examples below, as well as in thefigures, are intended to encompass and include all variations andisomers of the structure which are foreseeable, including allstereoisomers and constitutional or configurational isomers when theillustration, description, or figure is not explicitly limited to anyparticular isomer.

Methods for Preparing Cyclic Synthons

To avoid the need to separate single configurational or enantiomericisomers from complex mixtures resulting from non-specific reactions,stereospecific or at least stereoselective coupling reactions may beemployed in the preparation of the synthons of this invention. Thefollowing are examples of synthetic schemes for several classes ofsynthons useful in the preparation of macrocyclic modules of thisinvention. In general, the core synthons are illustrated, and lipophilicmoieties are not shown on the structures, however, it is understood thatall of the following synthetic schemes might encompass additionallipophilic or hydrophilic moieties used to prepare amphiphilic and othermodified macrocyclic modules. Species are numbered in relation to thescheme in which they appear; for example, “S1-1” refers to the structure1 in Scheme 1.

An approach to preparing synthons of 1,3-Diaminocyclohex-5-ene is shownin Scheme 1. Enzymatically assisted partial hydrolysis of the

symmetrical diester S1-1 is used to give enantiomerically pure S1-2.S1-2 is subjected to the Curtius reaction and then quenched with benzylalcohol to give protected amino acid S1-3. Iodolactonization ofcarboxylic acid S1-4 followed by dehydrohalogenation gives unsaturatedlactone S1-6. Opening of the lactone ring with sodium methoxide givesalcohol S1-7, which is converted with inversion of configuration to S1-8in a one-pot reaction involving mesylation, SN₂ displacement with azide,reduction and protection of the resulting amine with di-tert-butyldicarbonate. Epimerization of S1-8 to the more stable diequatorialconfiguration followed by saponification gives carboxylic acid S1-10.S1-10 is subjected to the Curtius reaction. A mixed anhydride isprepared using ethyl chloroformate followed by reaction with aqueousNaN₃ to give the acyl azide, which is thermally rearranged to theisocyanate in refluxing benzene. The isocyanate is quenched with2-trimethylsilylethanol to give differentially protected tricarbamateS1-11. Reaction with trifluoroacetic acid (TFA) selectively deprotectsthe 1,3-diamino groups to provide the desired synthon S1-12.

In another variation, an approach to preparing synthons of1,3-Diaminocyclohexane is shown in Scheme 1a.

Some aspects of these preparations are given in Suami et al., J. Org.Chem. 1975, 40, 456 and Kavadias et al. Can. J. Chem. 1978, 56, 404.

In another variation, an approach to preparing synthons of1,3-substituted cyclohexane is shown in Scheme 1b.

This synthon will remain “Z-protected” until the macrocyclic module hasbeen cyclized. Subsequent deprotection to yield a macrocyclic modulewith amine functional groups is done by a hydrogenation protocol.

Norbornanes (bicycloheptanes) may be used to prepare synthons of thisinvention, and stereochemically controlled multifunctionalization ofnorbornanes can be achieved. For example, Diels-Alder cycloaddition maybe used to form norbornanes incorporating various functional groupshaving specific, predictable stereochemistry. Enantiomerically enhancedproducts may also be obtained through the use of appropriate reagents,thus limiting the need for chiral separations.

An approach to preparing synthons of 1,2-Diaminonorbornane is shown inScheme 2. 5-(Benzyloxy-methyl)-1,3-cyclopentadiene (S2-13) is reactedwith

diethylaluminum chloride Lewis acid complex of di-(l)-menthyl fumarate(S2-14) at low temperature to give the diastereomerically purenorbornene S2-15. Saponification with potassium hydroxide in aqueousethanol gives the diacid S2-16, which is subjected to a tandem Curtiusreaction with diphenylphosphoryl azide (DPPA), the reaction product isquenched with 2-trimethylsilylethanol to give the biscarbamate S2-17.Deprotection with TFA gives diamine S2-18.

Another approach to this synthon class is outlined in Scheme 3. Openingof anhydride S3-19 with methanol in the presence of quinidine gives theenantiomerically pure ester acid S3-20. Epimerization of the ester groupwith sodium methoxide (NaOMe) gives S3-21. A Curtius reaction with DPPAfollowed by quenching with trimethylsilylethanol gives carbamate S3-22.Saponification with NaOH gives the acid S3-23, which undergoes a Curtiusreaction,

then quenched with benzyl alcohol to give differentially protectedbiscarbamate S3-24. Compound S3-24 can be fully deprotected to providethe diamine or either of the carbamates can be selectively deprotected.

An approach to preparing synthons of endo,endo-1,3-Diaminonorbornane isshown in Scheme 4. 5-Trimethylsilyl-1,3-cyclopentadiene (S4-25) isreacted with the diethylaluminum chloride Lewis acid complex ofdi-(l)-menthyl fumarate at low temperature to give nearlydiastereomerically pure norbornene S4-26. Crystallization of S4-26 fromalcohol results in recovery of greater than 99% of the singlediastereomer. Bromolactonization followed by silver mediatedrearrangement gives mixed diester S4-28 with an alcohol moiety at the7-position. Protection of the alcohol with benzyl bromide and selectivedeprotection of the methyl ester gives the free carboxylic acid S4-30. ACurtius reaction results in trimethylsilylethyl carbamate norborneneS4-31. Biscarbonylation of the olefin in methanol, followed by asingle-step deprotection and dehydration gives the mono-anhydride S4-33.Quinidine mediated opening of the anhydride with methanol gives S4-34.Curtius transformation of S4-34 gives the biscarbamate S4-35, which isdeprotected with TFA or tetrabutylammonium fluoride (TBAF) to givediamine S4-36.

Another approach to this class of synthons is outlined in Scheme 5.Benzyl alcohol opening of S3-19 in the presence of quinidine gives S5-37in high enantiomeric excess. Iodolactonization followed by NaBT₄reduction gives lactone S5-39. Treatment with NaOMe liberates the methylester and the free alcohol to generate S5-40. Transformation of thealcohol S5-40 to the inverted t-butyl carbamate protected amine S5-41 isaccomplished in a one-pot reaction by azide deplacement of the mesylateS5-40 followed by reduction to the amine, which is protected withdi-tert-butyl dicarbonate. Hydrogenolytic cleavage of the benzyl esterand epimerization of the methyl ester to the exo configuration isfollowed by protection of the free acid with benzyl bromide to giveS5-44. Saponification of the methyl ester followed by atrimethylsilylethanol quenched Curtius reaction

gives the biscarbamate S5-46, which is cleaved with TFA to give thedesired diamine S5-47.

An approach to preparing synthons of exo,endo-1,3-Diaminonorbornane isshown in Scheme 6. p-Methoxybenzyl alcohol opening of norborneneanhydride S3-19 in the presence of quinidine gives monoester S6-48 inhigh enantiomeric excess. Curtius reaction of the free acid givesprotected all endo monoacid-monoamine S6-49. Biscarbonylation andanhydride formation gives exo-monoanhydride S6-51. Selectivemethanolysis in the presence of quinine gives S6-52. Atrimethylsilylethanol quenched Curtius reaction gives biscarbamateS6-53. Epimerization of the two esters results in the more stericallystable S6-54. Cleavage of the carbamate groups provides synthon S6-55.

Methods to Prepare Macrocyclic Modules

Synthons may be coupled to one another to form macrocyclic modules. Inone variation, the coupling of synthons may be accomplished in aconcerted scheme. Preparation of a macrocyclic module by the concertedroute may be performed using, for example, at least two types ofsynthons, each type having at least two functional groups for couplingto other synthons. The functional groups may be selected so that afunctional group of one type of synthon can couple only to a functionalgroup of the other type of synthon. When two types of synthons are used,a macrocyclic module may be formed having alternating synthons ofdifferent types. Scheme 7 illustrates a concerted module synthesis.

Referring to Scheme 7, 1,2-Diaminocyclohexane, S7-1, is a synthon havingtwo amino functional groups for coupling to other synthons, and2,6-diformyl-4-dodec-1-ynylphenol, S7-2, is a synthon having two formylgroups for coupling to other synthons. An amino group may couple with aformyl group to form an imine linkage. In Scheme 7, a concerted producthexamer macrocyclic module is shown.

In one variation, a mixture of tetramer, hexamer, and octamermacrocyclic modules may be formed in the concerted scheme. The yields ofthese macrocyclic modules can be varied by changing the concentration ofvarious synthons in the reagent mixture, and among other factors, bychanging the solvent, temperature, and reaction time.

The imine groups of S7-3 can be reduced, e.g. with sodium borohydride,to give amine linkages. If the reaction is carried out using2,6-di(chlorocarbonyl)-4-dodec-1-ynylphenol instead of2,6-diformyl-4-dodec-1-ynylphenol, the resulting module will containamide linkages. Similarly, if 1,2-dihydroxycyclohexane is reacted with2,6-di(chlorocarbonyl)-4-dodec-1-ynylphenol, the resulting module willcontain ester linkages.

In some variations, the coupling of synthons may be accomplished in astepwise scheme. In an example of the stepwise preparation ofmacrocyclic modules, a first type of synthon is substituted with oneprotected functional group and one unprotected functional group. Asecond type of synthon is substituted with an unprotected functionalgroup that will couple with the unprotected functional group on thefirst synthon. The product of contacting the first type of synthon withthe second type of synthon may be a dimer, which is made of two coupledsynthons. The second synthon may also be substituted with anotherfunctional group which is either protected, or which does not couplewith the first synthon when the dimer is formed. The dimer may beisolated and purified, or the preparation may proceed as a one-potmethod. The dimer may be contacted with a third synthon having twofunctional groups, only one of which may couple with the remainingfunctional group of either the first or second synthons to form atrimer, which is made of three coupled synthons. Such stepwise couplingof synthons may be repeated to form macrocyclic modules of various ringsizes. To cyclize or close the ring of the macrocyclic module, then^(th) synthon which was coupled to the product may be substituted witha second functional group which may couple with the second functionalgroup of a previously coupled synthon that has not been coupled, whichmay be deprotected for that step. The stepwise method may be carried outwith synthons on solid phase support. Scheme 8 illustrates a stepwisepreparation of module SC8-1.

Compound S8-2 is reacted with S8-3, in which the phenol is protected asthe benzyl ether and the nitrogen is shown as protected with a group“P,” which can be any of a large number of protecting groups well-knownin the art, in the presence of methanesulfonyl chloride (Endo, K.;Takahashi, H. Heterocycles, 1999, 51, 337), to give S8-4. Removal of theN-protecting group give the free amine S8-5, which can be coupled withsynthon S8-6 using any standard peptide coupling reaction such asBOP/HOBt to give S8-7. Deprotection/coupling is repeated, alternatingsynthons S8-3 and S8-6 until a linear construct with eight residues isobtained. The remaining acid and amine protecting groups on the 8-merare removed and the oligomer is cyclized, see e.g., Caba, J. M., et al.,J. Org. Chem., 2001, 66:7568 (PyAOP cyclization) and Tarver, J. E. etal., J. Org. Chem., 2001, 66:7575 (active ester cyclization). The Rgroup is H or an alkyl group linked via a functional group to thebenzene ring, and X is N, O, or S. Examples of solid supports includeWang resin, hydrogels, silica gels, sepharose, sephadex, agarose, andinorganic solids. Using a solid support might simplify the procedure byobviating purification of intermediates along the way. The finalcyclization may be done in a solid phase mode. A “safety-catch linker”approach (Bourne, G. T., et al., J. Org. Chem., 2001, 66:7706) may beused to obtain cyclization and resin cleavage in a single operation.

In another variation, a concerted method involves contacting two or moredifferent synthons and a linker molecule as shown in Scheme 9, where Rmay be an alkyl group or other lipophilic group.

In another variation, a stepwise linear method involves various synthonsand a solid phase support as shown in Scheme 10.

In another variation, a stepwise convergent method involves synthontrimers and a solid phase support as shown in Scheme 11. This method canalso be done without the solid phase support using trimers in solution.

In another variation, a template method involves synthons broughttogether by a template as shown in Scheme 12. Some aspects of thisapproach (and an Mg²⁺ template) are given in Dutta et al. Inorg. Chem.1998, 37, 5029.

In another variation, a linker molecule method involves cyclizingsynthons in solution as shown in Scheme 13.

Reagents for the following examples were obtained from Aldrich ChemicalCompany and VWR Scientific Products. All reactions were carried outunder nitrogen or argon atmosphere unless otherwise noted. Solventextracts of aqueous solutions were dried over anhydrous Na₂SO₄.Solutions were concentrated under reduced pressure using a rotaryevaporator. Thin layer chromatography (TLC) was done on Analtech Silicagel GF (0.25 mm) plates or on Machery-Nagel Alugram Sil G/UV (0.20 mm)plates. Chromatograms were visualized with either UV light,phosphomolybdic acid, or KMnO₄. All compounds reported were homogenousby TLC unless otherwise noted. HPLC analyses were performed on a HewlettPackard 1100 system using a reverse phase C-18 silica column.Enantiomeric excess was determined by HPLC using a reverse phase(l)-leucine silica column from Regis Technologies. All ¹[H] and ¹³[C]NMR spectra were collected at 400 MHz on a Varian Mercury system.Electrospray mass spectra were obtained by Synpep Corp., or on a ThermoFinnigan LC-MS system.

Example 16 2,6-Diformyl-4-bromophenol

Hexamethylenetetramine (73.84 g, 526 mmol) was added to TFA (240 mL)with stirring. 4-Bromophenol (22.74 g, 131 mmol) was added in oneportion and the solution heated in an oil bath to 120° C. and stirredunder argon for 48 h. The reaction mixture was then cooled to ambienttemperature. Water (160 mL) and 50% aqueous H₂SO₄ (80 mL) were added andthe solution stirred for an additional 2 h. The reaction mixture waspoured into water (1600 mL) and the resulting precipitate collected on aBüchner funnel. The precipitate was dissolved in ethyl acetate (EtOAc)and the solution was dried over MgSO₄. The solution was filtered and thesolvent removed on a rotary evaporator. Purification by columnchromatography on silica gel (400 g) using a gradient of 15-40% ethylacetate in hexanes resulted in a isolation of the product as a yellowsolid (18.0 g, 60%).

¹H NMR (400 MHz, CDCl₃) δ 11.54 (s, 1H, OH), 10.19 (s, 2H, CHO), 8.08(s, 2H, ArH).

Example 17 2,6-Diformyl-4-(dodecyn-1-yl)phenol

2,6-Diformyl-4-bromophenol (2.50 g, 10.9 mmol), 1-dodecyne (2.00 g, 12.0mmol), CuI (65 mg, 0.33 mmol), and bis(triphenylphosphine)palladium)II)dichloride were suspended in degassed acetonitrile (MeCN) (5 mL) anddegassed benzene (1 mL). The yellow suspension was sparged with argonfor 30 min and degassed Et₃N (1 mL) was added. The resulting brownsuspension was sealed in a pressure vial, warmed to 80° C. and heldthere for 12 h. The mixture was then partitioned between EtOAc and KHSO₄solution. The organic layer was separated, washed with brine, dried(MgSO₄) and concentrated under reduced pressure. The dark yellow oil waspurified by column chromatography on silica gel (25% Et₂O in hexanes) togive 1.56 g (46%) of the title compound.

¹H NMR (400 MHz, CDCl₃) δ11.64 (s, 1H, OH), 10.19 (s, 2H, CHO), 7.97 (s,2H, ArH), 2.39 (t, 2H, J=7.2 Hz, propargylic), 1.59 (m, 3H, aliphatic),1.43, (m, 2H, aliphatic), 1.28 (m, 11H, aliphatic), 0.88 (t, 3H, J=7.0Hz, CH₃).

¹³C NMR (400 MHz, CDCl₃) δ 192.5, 162.4, 140.3, 122.8, 116.7, 91.4,77.5, 31.9, 29.6, 29.5, 29.3, 29.1, 28.9, 28.5, 22.7, 19.2, 14.1.

MS (FAB): Calcd. for C₂₀H₂₇O₃, 315.1960. found 315.1958 [M+H]⁺.

Example 18 2,6-Diformyl-4-(dodecen-1-yl)phenol

2,6-Diformyl-4-bromophenol (1.00 g, 4.37 mmol), 1-dodecene (4.8 mL, 21.7mmol), 1.40 g tetrabutylammonium bromide (4.34 mmol), 0.50 g NaHCO₃(5.95 mmol), 1.00 g LiCl (23.6 mmol) and 0.100 g palladium diacetate(Pd(OAc)₂) (0.45 mmol) were combined in 30 mL degassed anhydrousdimethylformamide (DMF). The mixture was sparged with argon for 10 minand then sealed in a pressure vial which was warmed to 82° C. and heldfor 40 h. The crude reaction mixture was partitioned between CH₂Cl₂ and0.1 M HCl solution. The organic layer was washed with 0.1 M HCl (2×),brine (2×), and saturated aqueous NaHCO₃ (2×), dried over MgSO₄ andconcentrated under reduced pressure. The dark yellow oil was purified bycolumn chromatography on silica gel (25% hexanes in Et₂O) to give 0.700g (51%) of the title compound as primarily the Z isomer.

¹H NMR (400 MHz, CDCl₃) δ11.50 (s, 1H, OH), 10.21 (s, 2H, CHO), 7.95 (s,2H, ArH), 6.38 (d, 1H, vinyl), 6.25 (m, 1H, vinyl), 2.21 (m, 2H,allylic), 1.30-1.61 (m, 16H, aliphatic), 0.95 (t, 3H, J=7.0 Hz, CH₃).

MS (FAB): Calcd. for C₂₀H₂₇O₃, 315.20. found 315.35 [M−H]⁻.

Example 19 (1R,6S)-6-Methoxycarbonyl-3-cyclohexene-1-carboxylic Acid(S1-2)

S1-1 (15.0 g, 75.7 mmol) was suspended in pH 7 phosphate buffer (950mL). Pig liver esterase (2909 units) was added, and the mixture stirredat ambient temperature for 72 h with the pH maintained at 7 by additionof 2M NaOH. The reaction mixture was washed with ethyl acetate (200 mL),acidified to pH 2 with 2M HCl, and extracted with ethyl acetate (3×200mL). The extracts were combined, dried, and evaporated to afford 13.8 g(99%) of S1-2.

¹H NMR: (CDCl₃) δ 2.32 (dt, 2H, 2_(ax)- and 5_(ax)-H's), 2.55 (dt, 2H,2_(eq)- and 5_(eq)-H's), 3.00 (m, 2H, 1- and 6-H's), 3.62 (s, 3H,CO₂Me), 5.61 (m, 2H, 3- and 4-H's).

Example 20 Methyl(1S,6R)-6-Benzyloxycarbonylaminocyclohex-3-enecarboxylate (S1-3)

S1-2 (10.0 g, 54.3 mmol) was dissolved in benzene (100 mL) under N₂.Triethylamine (13.2 g, 18.2 mL, 130.3 mmol) was added followed by DPPA(14.9 g, 11.7 mL, 54.3 mmol). The solution was refluxed for 20 h. Benzylalcohol (5.9 g, 5.6 mL, 54.3 mmol) was added and reflux continued for 20h. The solution was diluted with EtOAc (200 mL), washed with saturatedaqueous NaHCO₃ (2×50 mL), water (20 mL), and saturated aqueous NaCl (20mL), dried and evaporated to give 13.7 g (87%) of S1-3.

¹H NMR: (CDCl₃) δ 2.19 (dt, 1H, 5_(ax)-H), 2.37 (tt, 2H, 2_(ax)- and5_(eq)-H's), 2.54 (dt, 1H, 2_(eq)-H), 2.82 (m, 1H, 1-H), 3.65 (s, 3H,CO₂Me), 4.28 (m, 1H, 6-H), 5.08 (dd, 2H, CH₂Ar), 5.42 (d, 1H, NH), 5.62(ddt, 2H, 3- and 4-H's), 7.35 (m, 5H, Ar H's).

Example 21 (1S,6R)-6-Benzyloxycarbonylaminocyclohex-3-enecarboxylic acid(S1-4)

S1-3 (23.5 g, 81.3 mmol) was dissolved in MeOH (150 mL) and the solutioncooled to 0° C. 2M NaOH (204 mL, 0.41 mol) was added, the mixtureallowed to come to ambient temperature and then it was stirred for 48 h.The reaction mixture was diluted with water (300 mL), acidified with 2MHCl, and extracted with dichloromethane (250 mL), dried, and evaporated.The residue was recrystallized from diethyl ether to give 21.7 (97%) ofS1-4.

¹H NMR: (CDCl₃) δ 2.20 (d, 1H, 5_(ax)-H), 2.37 (d, 2H, 2_(ax)- and5_(eq)-H's), 2.54 (d, 1H, 2_(eq)-H), 2.90 (br s, 1H, 1-H), 4.24 (br s,1H, 6-H), 5.08 (dd, 2H, CH₂Ar), 5.48 (d, 1H, NH), 5.62 (dd, 2H, 3- and4-H's), 7.35 (m, 5H, Ar H's).

Example 22(1S,2R,4R,5R)-2-Benzyloxycarbonylamino-4-iodo-7-oxo-6-oxabicyclo[3.2.1]octane(S1-5)

S1-4 (13.9 g, 50.5 mmol) was dissolved in dichloromethane (100 mL) underN₂, 0.5 M NaHCO₃ (300 mL), KI (50.3 g, 303.3 mmol), and iodine (25.6 g,101 mmol) were added and the mixture stirred at ambient temperature for72 h. The mixture was diluted with dichloromethane (50 mL) and theorganic phase separated. The organic phase was washed with saturatedaqueous Na₂S₂O₃ (2×50 mL), water (30 mL), and saturated aqueous NaCl (20mL), dried and evaporated to afford 16.3 g (80%) of S1-5.

¹H NMR: (CDCl₃) δ 2.15 (m, 1H, 8_(ax)-H), 2.42 (m, 2H, 3_(ax)- and8_(eq)-H's), 2.75 (m, 2H, 1- and 3_(eq)-H's), 4.12 (br s, 1H, 2-H), 4.41(t, 1H, 4-H), 4.76 (dd, 1H, 5-H), 4.92 (d, 1H, NH), 5.08 (dd, 2H,CH₂Ar), 7.35 (m, 5H, Ar H's).

Example 23(1S,2R,5R)-2-Benzyloxycarbonylamino-7-oxo-6-oxabicyclo[3.2.1]oct-3-ene(S1-6)

S1-5 (4.0 g, 10 mmol) was dissolved in benzene (50 mL) under N₂.1,8-Diazabicyclo[5.4.0]undec-7-ene (DBU) (1.8 g, 12 mmol) was added andthe solution refluxed for 16 h. The precipitate was filtered and thefiltrate was diluted with EtOAc (200 mL). The filtrate was washed with1M HCl (20 mL), saturated aqueous Na₂S₂O₃ (20 mL), water (20 mL), andsaturated aqueous NaCl (20 mL), dried and evaporated to give 2.2 g (81%)S1-6.

¹H NMR: (CDCl₃) δ 2.18 (d, 1H, 8_(ax)-H), 2.39 (m, 1H, 8_(eq)-H), 3.04(t, 1H, 1-H), 4.70 (m, 1H, 5-H), 4.82 (t, 1H, 2-H), 5.15 (dd, 3H, CH₂Arand NH), 5.76 (d, 1H, 4-H), 5.92 (m, 1H, 3-H), 7.36 (s, 5H, Ar H's).

Example 24 (1S,2R,5R)-Methyl2-Benzyloxycarbonylamino-5-hydroxycyclohex-3-enecarboxylate (S1-7)

S1-6 (9.0 g, 33 mmol) was suspended in MeOH (90 mL) and cooled to 0° C.NaOMe (2.8 g, 52.7 mmol) was added and the mixture stirred for 3 hduring which time a solution gradually formed. The solution wasneutralized with 2M HCl, diluted with saturated aqueous NaCl (200 mL),and extracted with dichloromethane (2×100 mL). The extracts werecombined, washed with water (20 mL) and saturated aqueous NaCl (20 ml),dried, and evaporated. The residue was flash chromatographed (silica gel(250 g), 50:50 hexane/EtOAc) to give 8.5 g (85%) of S1-7.

¹H NMR: (CDCl₃) δ 1.90 (m, 1H, 6_(ax)-H), 2.09 (m, 1H, 6_(eq)-H), 2.81(m, 1H, 1-H), 3.55 (s, 3H, CO₂Me), 4.15 (m, 1H, 5-H), 4.48 (t, 1H, 2-H),5.02 (dd, 2H, CH₂Ar), 5.32 (d, 1H, NH), 5.64 (dt, 1H, 4-H), 5.82 (dt,1H, 3-H), 7.28 (s, 5H, Ar H's).

Example 25 (1S,2R,5S)-Methyl2-Benzyloxycarbonylamino-5-t-butoxycarbonylaminocyclohex-3-enecarboxylate(S1-8)

S1-7 (7.9 g, 25.9 mmol) was dissolved in dichloromethane (150 mL) andcooled to 0° C. under N₂. Triethylamine (6.3 g, 8.7 mL, 62.1 mmol) andmethanesulfonyl chloride (7.1 g, 62.1 mmol) were added and the mixturestirred at 0° C. for 2 h. (n-Bu)₄NN₃ (14.7 g, 51.7 mmol) indichloromethane (50 mL) was added and stirring continued at 0° C. for 3h followed by 15 h at ambient temperature. The mixture was cooled to 0°C. and P(n-Bu)₃ (15.7 g, 19.3 mL, 77.7 mmol) and water (1 mL) were addedand the mixture stirred at ambient temperature for 24 h. Di-tert-butyldicarbonate (17.0 g, 77.7 mmol) was added and stirring continued for 24h. The solvent was removed, the residue dissolved in 2:1 hexane/EtOAc(100 mL), the solution filtered, and evaporated. The residue was flashchromatographed (silica gel (240 g), 67:33 hexane/EtOAc) to give 5.9 g(56%) of S1-8.

¹H NMR: (CDCl₃) δ 1.40 (s, 9H, Boc H's), 1.88 (m, 1H, 6_(ax)-H), 2.21(m, 1H, 6_(eq)-H), 2.95 (m, 1H, 1-H), 3.60 (s, 3H, CO₂Me), 4.15 (d, 1H,Boc NH), 4.50 (m, 2H, 2- and 5-H's), 5.02 (s, 2H, CH₂Ar), 5.38 (d, 1H, ZNH), 5.65 (m, 2 H, 3- and 4-H's), 7.30 (s, 5H, Ar H's).

Example 26 (1R,2R,3S)-Methyl2-Benzyloxycarbonylamino-5-t-butoxycarbonylaminocyclohex-3-enecarboxylate(S1-9)

S1-8 (1.1 g, 2.7 mmol) was suspended in MeOH (50 mL). NaOMe (0.73 g,13.6 mmol) was added and the mixture refluxed for 18 h after which 0.5 MNH₄Cl (50 mL) was added and the resulting precipitate collected. Thefiltrate was evaporated and the residue triturated with water (25 mL).The insoluble portion was collected and combined with the originalprecipitate to give 0.85 g (77%) of S1-9.

¹H NMR: (CDCl₃) δ 1.38 (s, 9H, Boc H's), 1.66 (m, 1H, 6_(ax)-H), 2.22(d, 1H, 6_(eq)-H), 2.58 (t, 1H, 1-H), 3.59 (3, 3H, CO₂Me), 4.22 (br s,1H, Boc NH), 4.50 (m, 2H, 2- and 5-H's), 4.75 (d, 1H, Z NH), 5.02 (s,2H, CH₂Ar), 5.62 (s, 2H, 3- and 4-H's), 7.30 (s, 5H, Ar H's).

Example 27(1R,2R,5S)-2-benzyloxycarbonylamino-5-t-butoxycarbonylaminocyclohex-3-enecarboxylicacid (S1-10)

S1-9 (0.85 g, 2.1 mmol) was suspended in 50:50 MeOH/dichloromethane (5mL) and cooled to 0° C. under N₂ after which 2M NaOH (2.0 mL) was addedand the mixture stirred at ambient temperature for 16 h. The mixture wasacidified with 2M HCl upon which a white precipitate formed. Theprecipitate was collected, washed with water and hexane, and dried togive 0.74 g (90%) of S1-10.

¹H NMR: (CD₃OD) δ 1.42 (s, 9H, Boc H's), 1.66 (m, 1H, 6_(ax)-H), 2.22(d, 1H, 6_(eq)-H), 2.65 (t, 1H, 1-H), 4.18 (m, 1H, 5-H), 4.45 (m, 1H,5-H), 5.04 (s, 2H, CH₂Ar), 5.58 (m, 2H, 3- and 4-H's), 7.35 (s, 5H, ArH's).

Example 28(1R,2R,5S)-2-Benzyloxycarbonylamino-5-t-butoxycarbonylamino-1-(2-trimethylsilyl)ethoxycarbonylaminocyclohex-3-ene(S1-11)

S1-10 (3.1 g, 7.9 mmol) was dissolved in THF (30 mL) under N₂ and cooledto 0° C. Triethylamine (1.6 g, 2.2 mL, 15.9 mmol) was added followed byethyl chloroformate (1.3 g, 1.5 mL, 11.8 mmol). The mixture was stirredat 0° C. for 1 h. A solution of NaN₃ (1.3 g, 19.7 mmol) in water (10 mL)was added and stirring at 0° C. was continued for 2 h. The reactionmixture was partitioned between EtOAc (50 mL) and water (50 mL). Theorganic phase was separated, dried, and evaporated. The residue wasdissolved in benzene (50 mL) and refluxed for 2 h.2-Trimethylsilylethanol (1.0 g, 1.2 mL, 8.7 mmol) was added and refluxcontinued for 3 h. The reaction mixture was diluted with EtOAc (200 mL),washed with saturated aqueous NaHCO₃ (50 mL), water (20 mL), andsaturated aqueous NaCl (20 mL), dried and evaporated. The residue wasflash chromatographed (silica gel (100 g), 67:33 hexane/EtOAc) to give3.1 g (77%) of S1-11.

¹H NMR: (CDCl₃) δ −0.02 (s, 9H, TMS), 0.90 (t, 3H, CH₂TMS), 1.40 (s, 9H,Boc H's), 2.38 (m, 1H, 6_(eq)-H), 3.62 (m, 1H, 1-H), 4.08 (m, 2H,OCH₂CH₂TMS), 4.18 (m, 1H), 4.38 (m, 1H), 4.62 (m, 1H), 5.07 (dd, 2H,CH₂Ar), 5.18 (m, 1H), 5.26 (m, 1H), 5.58 (d, 1H, olefinic H), 5.64 (d,2H, olefinic H), 7.30 (s, 5, Ar H's).

Example 29 (1R,2R,5S)-2-Benzyloxycarbonylamino-1,5-diaminocyclohex-3-ene(S1-12)

S1-11 (2.5 g, 4.9 mmol) was added to TFA (10 mL) and the solutionstirred at ambient temperature for 16 h after which the solution wasevaporated. The residue was dissolved in water (20 mL), basified to pH14 with KOH and extracted with dichloromethane (3×50 mL). The extractswere combined, washed with water (20 mL), dried and evaporated to give1.1 g (85%) of S1-12.

¹H NMR: (CDCl₃) δ 1.30 (m, 1H, 61, —H), 2.15 (br d, 1H, 6_(eq)-H), 2.68(m, 1H, 1-H), 3.42 (br s, 1H, 5-H), 3.95 (m, 1H, 2-H), 4.85 (d, 1H, ZNH), 5.08 (t, 2H, CH₂Ar), 5.45 (d, 1H, 4-H), 5.62 (d, 1H, 3-H), 7.32 (s,5H, Ar H's). ESCI MS m/e 262 M+1.

Example 30

Isolation of S1b-2 was accomplished using the following procedure: UsingSchlenk technique 5.57 g (10.0 mmol) of methyl ester compound, S1b-1,was dissolved in 250 mL of THF. In another flask LiOH (1.21 g, 50.5mmol) was dissolved in 50 mL water and de-gassed by bubbling N₂ throughthe solution using a needle for 20 minutes. The reaction was startedtransferring the base solution into the flask containing S1b-1 over oneminute with rapid stirring. The mixture was stirred at room temperatureand work-up initiated when the starting material S1b-1 was completelyconsumed (Using a solvent system of 66% EtOAc/33% Hexane and developingwith phosphomolybdic acid reagent (Aldrich #31, 927-9) the startingmaterial S1b-1 has an Rf of 0.88 and the product streaks with an Rf ofapprox. 0.34 to 0.64.). The reaction usually takes 2 days. Work-Up: TheTHF was removed by vacuum transfer until about the same volume is leftas water added to the reaction, in this case 50 mL. During this thereaction solution forms a white mass that adheres to the stir barsurrounded by clear yellow solution. As the THF is being removed aseparatory funnel is set up including a funnel to pour in the reactionsolution and an Erlenmeyer flask is placed underneath the separatoryfunnel. Into the Erlenmeyer flask is added some anhydrous Na₂SO₄. Thisapparatus should be set up before acidification is started. (It isimportant to set up the separatory funnel and Erlenmeyer flask etc.before acidification of the reaction solution to enable separation ofphases and extraction of the product away from the acid quickly once thesolution attains a pH close to 1. If the separation is not preformedrapidly the Boc functional group will be hydrolyzed significantlyreducing the yield.) Once the volatiles are sufficiently removed, CH₂Cl₂(125 mL) and water (65 mL) are added and the reaction flask cooled in anice bath. The solution is stirred rapidly and 5 mL aliquots of 1N HClare added by syringe and the reaction solution tested with pH paper.Acid is added until the spot on the pH paper shows red (not orange)around the edge indicating a pH is 1 to 2 has been achieved (Thesolution being tested is a mixture of CH₂Cl₂ and water so the pH paperwill show the accurate measurement at the edge of the spot and not thecenter.) and the phases are separated by quickly pouring the solutioninto the separatory funnel. As the phases separate the stopcock isturned to release the CH₂Cl₂ phase (bottom) into the Erlenmeyer flaskand swirl the flask to allow the drying agent to absorb water in thesolution. (At this scale of this procedure 80 mL of 1N HCl was used.)Soon after phase separation the aqueous phase is extracted with CH₂Cl₂(2×100 mL), dried over anhydrous Na₂SO₄ and the volatiles removed toproduce 5.37 g/9.91 mmoles of a beautiful white microcrystals reflectinga 99.1% yield. This product can not be purified by chromatography sincethat process would also hydrolyze the Boc functional group on thecolumn.

¹H NMR (400 MHz, CDCl₃) δ7.33, 7.25 (5H, m, Ph), 6.30 (1H, d, NH), 5.97(1H, d, Nm), 5.10 (2H, m, CH₂Ph), 4.90 (1H, d, NH), 3.92, 3.58, 3.49(1H, m, CHNH), 2.96, 2.48, 2.04, 1.95, 1.63 (1H, m, CH₂CHNH), 1.34 (9H,s, CCH₃).

IR (crystalline, cm⁻¹) 3326 br w, 3066 w, 3033 w, 2975 w, 2940 w sh,1695 vs, 1506 vs, 1454 m sh, 1391 w, 1367 m, 1300 m sh, 1278 m sh, 1236s, 1213 w sh, 1163 vs, 1100 w, 1053 m, 1020 m, 981 w sh, 910 w, 870 m,846 w, 817 w, 775 w sh, 739 m, 696 m.

Example 31 Di-(l)-menthylbicyclo[2.2.1]hept-5-ene-7-anti-(trimethylsilyl)-2-endo-3-exo-dicarboxylate(S4-26)

To a solution of S4-25 (6.09 g, 0.0155 mol) in toluene (100 mL) wasadded diethylaluminum chloride (8.6 mL of a 1.8 M solution in toluene)at −78° C. under nitrogen and the mixture was stirred for 1 hour. To theresulting orange solution was added S2-14 (7.00 g, 0.0466 mol) dropwiseas a −78° C. solution in toluene (10 mL). The solution was kept at −78°C. for 2 hours, followed by slow warming to room temperature overnight.The aluminum reagent was quenched with a saturated solution of ammoniumchloride (50 mL). The aqueous layer was separated and extracted withmethylene chloride (100 mL) which was subsequently dried over magnesiumsulfate. Evaporation of the solvent left a yellow solid that waspurified by column chromatography (10% ethyl acetate/hexanes) to giveS4-26 as a while solid (7.19 g, 0.0136 mol, 87% yield).

¹H NMR: (CDCl₃) δ −0.09 (s, 9H, SiMe₃), 0.74-1.95 (multiplets, 36H,menthol), 2.72 (d, 1H, α-menthyl carbonyl CH), 3.19 (bs, 1H, bridgeheadCH), 3.30 (bs, 1H, bridgehead CH), 3.40 (t, 1H, α-menthyl carbonyl CH),4.48 (d of t, 1H, α-menthyl ester CH), 4.71 (d of t, 1H, α-menthyl esterCH), 5.92 (d of d, 1H, CH═CH), 6.19 (d of d, 1H, CH═CH).

Example 325-exo-Bromo-3-exo-1)-menthylcarboxybicyclo[2.2.1]heptane-7-anti-(trimethylsilyl)-2,6-carbolactone(S4-27)

A solution of bromine (3.61 g, 0.0226 mol) in methylene chloride (20 mL)was added to a stirring solution of S4-26 (4.00 g, 0.00754 mol) inmethylene chloride (80 mL). Stirring was continued at room temperatureovernight. The solution was treated with 5% sodium thiosulfate (150 mL),and the organic layer separated and dried over magnesium sulfate. Thesolvent was evaporated at reduced pressure, and the crude productpurified by column chromatography (5% ethyl acetate/hexanes) to giveS4-27 as a white solid (3.53 g, 0.00754 mol, 99% yield).

¹H NMR: (CDCl₃) δ −0.19 (s, 9H, SiMe₃), 0.74-1.91 (multiplets, 18H,menthol), 2.82 (d, 1H, α-lactone carbonyl CH), 3.14 (bs, 1H, lactonebridgehead CH), 3.19 (d of d, 1H, bridgehead CH), 3.29 (t, 1H, α-menthylcarbonyl CH), 3.80 (d, 1H, α-lactone ester), 4.74 (d of t, 1H, α-menthylester CH), 4.94 (d, 1H, bromo CH).

Example 33Bicyclo[2.2.1]hept-5-ene-7-syn-(hydroxy)-2-exo-methyl-3-endo-(l)-menthyldicarboxylate (S4-28)

S4-27 (3.00 g, 0.00638 mol) was dissolved in anhydrous methanol (150mL), silver nitrate (5.40 g, 0.0318 mol) added and the suspensionrefluxed for 3 days. The mixture was cooled, filtered through Celite andthe solvent evaporated to give an oily residue. Purification by columnchromatography gave S4-28 as a light yellow oil (1.72 g, 0.00491 mol,77% yield).

¹H NMR: (CDCl₃) δ 0.75-2.02 (multiplets, 18H, menthol), 2.83 (d, 1 H,α-menthyl carbonyl CH), 3.03 (bs, 1H, bridgehead CH), 3.14 (bs, 1H,bridgehead CH), 3.53 (t, 1H, α-methyl carbonyl CH), 3.76 (s, 3H, CH₃),4.62 (d of t, 1H, α-menthyl ester CH), 5.87 (d of d, 1H, CH═CH), 6.23 (dof d, 1H, CH═CH).

Example 342-exo-Methyl-3-endo-(l)-menthylbicyclo[2.2.1]hept-5-ene-7-syn-(benzyloxy)dicarboxylate(S4-29)

Benzyl bromide (1.20 g, 0.0070 mol) and silver oxide (1.62 g, 0.0070mol) were added to a stirring solution of S4-28 (0.490 g, 0.00140 mol)in DMF (25 mL). The suspension was stirred overnight and then dilutedwith ethyl acetate (100 mL). The solution was washed repeatedly withwater followed by 1 N lithium chloride. The organic layer was separatedand dried with magnesium sulfate. The solvent was evaporated underreduced pressure and the crude product was purified by columnchromatography on silica gel to give S4-29 as an oil (0.220 g, 0.000500mol, 36% yield).

¹H NMR: (CDCl₃) δ 0.74-2.08 (multiplets, 18H, menthol), 2.83 (d, 1 H,α-menthyl carbonyl CH), 3.18 (bs, 1H, bridgehead CH), 3.44 (bs, 1H,bridgehead CH), 3.52 (t, 1H, bridge CH), 3.57 (s, 3H, CH₃), 3.68 (t, 1H,α-methyl carbonyl CH), 4.42 (d of d, 2H, benzyl —CH₂—), 4.61 (d of t,1H, α-menthyl ester CH), 5.89 (d of d, 1H, CH═CH), 6.22 (d of d, 1H,CH═CH), 7.25-7.38 (m, 5H, C₆H₅).

Example 35Bicyclo[2.2.1]hept-5-ene-7-syn-(benzyloxy)-2-exo-carboxy-3-endo-(l)-menthylcarboxylate (S4-30)

S4-29 (0.220 g, 0.00050 mol) was added to a mixture of tetrahydrofuran(1.5 mL), water (0.5 mL), and methanol (0.5 mL). Potassium hydroxide(0.036 g, 0.00065 mol) was added and the solution stirred at roomtemperature overnight. The solvent was evaporated under reduced pressureand the residue purified by column chromatography (10% ethylacetate/hexanes) to give S4-30 (0.050 g, 0.00012 mol, 23% yield).

¹H NMR: (CDCl₃) δ 0.73-2.01 (multiplets, 18H, menthol), 2.85 (d, 1 H,α-menthyl carbonyl CH), 3.18 (bs, 1H, bridgehead CH), 3.98 (bs, 1H,bridgehead CH), 3.53 (bs, 1H, bridge CH), 3.66 (t, 1H, α-methyl carbonylCH), 4.44 (d of d, 2H, benzyl —CH₂—), 4.63 (d of t, 1H, α-menthyl esterCH), 5.90 (d of d, 1H, CH═CH), 6.23 (d of d, 1H, CH═CH), 7.25-7.38 (m,5H, C₆H₅).

Mass Spec: calculated for C₂₆H₃₄O₅, 426.24. found 425.4 (M−1) and 851.3(2M−1).

Example 36Bicyclo[2.2.1]hept-5-ene-7-syn-(benzyloxy)-2-exo-(trimethylsilylethoxycarbonyl)-amino-3-endo-(l)-menthylcarboxylate (S4-31)

To a solution of S4-30 in benzene is added triethylamine anddiphenylphosphoryl azide. The solution is refluxed for 24 hours thencooled to room temperature. Trimethylsilylethanol is added, and thesolution refluxed for an additional 48 hours. The benzene solution ispartitioned between ethyl acetate and 1 M sodium bicarbonate. Theorganic layers are combined, washed with 1 M sodium bicarbonate anddried over sodium sulfate. The solvent is evaporated under reducedpressure to give the crude Curtius reaction product.

Example 37Bicyclo[2.2.1]heptane-7-syn-(benzyloxy)-2-exo-(trimethylsilylethoxycarbonyl)-amino-3-endo-(l)-menthyl-5-exo-methyl-6-exo-methyltricarboxylate (S4-32)

S4-31, dry copper(II) chloride, 10% Pd/C, and dry methanol are added toa flask with vigorous stirring. After degassing, the flask is chargedwith carbon monoxide to a pressure just above 1 atm., which ismaintained for 72 hours. The solids are filtered and the residue workedup in the usual way to afford the biscarbonylation product.

Example 38Bicyclo[2.2.1]heptane-7-syn-(benzyloxy)-2-exo-(trimethylsilylethoxycarbonyl)-amino-3-endo-(l)-menthylcarbox-5-exo-6-exo-dicarboxylicanhydride (S4-33)

A mixture of S4-32, formic acid, and a catalytic amount ofp-toluenesulfonic acid is stirred at 90° C. overnight. Acetic anhydrideis added and the reaction mixture refluxed for 6 hours. Removal of thesolvents and washing with ether gives the desired anhydride.

Example 39Bicyclo[2.2.1]heptane-7-syn-(benzyloxy)-2-exo-(trimethylsilylethoxycarbonyl)-amino-3-endo-(l)-menthyl-6-exo-carboxy-5-exo-methyldicarboxylate (S4-33)

To a solution of S4-32 in equal amounts of toluene and carbontetrachloride is added quinidine. The suspension is cooled to −65° C.and stirred for 1 hour. Three equivalents of methanol are slowly addedover 30 minutes. The suspension is stirred at −65° C. for 4 daysfollowed by removal of the solvents under reduced pressure. Theresulting white solid is partitioned between ethyl acetate and 2M HCl.The quinine is recovered from the acid layer and S4-33 obtained from theorganic layer.

Example 40Bicyclo[2.2.1]heptane-7-syn-(benzyloxy)-2-exo-(trimethylsilylethoxycarbonyl)-amino-3-endo-(l)-menthyl-6-exo-(trimethylsilylethoxycarbonyl)amino-5-exo-methyldicarboxylate (S4-35)

To a solution of S4-34 in benzene is added triethylamine anddiphenylphosphoryl azide. The solution is refluxed for 24 hours. Aftercooling to room temperature, 2-trimethylsilylethanol is added and thesolution refluxed for 48 hours. The benzene solution is partitionedbetween ethyl acetate and 1M sodium bicarbonate. The organic layers arecombined, washed with 1M sodium bicarbonate, and dried over sodiumsulfate. The solvent is evaporated under reduced pressure to give thecrude Curtius reaction product.

Example 41endo-Bicyclo[2.2.1]hept-5-ene-2-benzylcarboxylate-3-carboxylic acid(S5-37)

Compound S3-19 (4.00 g, 0.0244 mol) and quinidine (8.63 g, 0.0266 mol)were suspended in equal amounts of toluene (50 mL) and carbontetrachloride (50 mL). The suspension was cooled to −55° C. after whichbenzyl alcohol (7.90 g, 0.0732 mol) was added over 15 minutes. Thereaction mixture became homogenous after 3 hours and was stirred at −55°C. for an additional 96 hours. After removal of the solvents, theresidue was partitioned between ethyl acetate (300 mL) and 2Mhydrochloric acid (100 mL). The organic layer was washed with water(2×50 mL) and saturated aqueous sodium chloride (1×50 mL). Drying overmagnesium sulfate and evaporation of the solvent gave S5-37 (4.17 g,0.0153 mol, 63% yield).

¹H NMR: (CDCl₃) δ 1.33 (d, 1H, bridge CH₂), 1.48 (d of t, 1H, bridgeCH₂), 3.18 (bs, 1H, bridgehead CH), 3.21 (bs, 1H, bridgehead CH), 3.33(t, 2H, α-acid CH), 4.98 (d of d, 2H, CH₂Ph), 6.22 (d of d, 1H, CH═CH),6.29 (d of d, 1H, CH═CH), 7.30 (m, 5H, C₆H₅).

Example 422-endo-Benzylcarboxy-6-exo-iodobicyclo[2.2.1]heptane-3,5-carbolactone(S5-38)

S5-37 (4.10 g, 0.0151 mol) was dissolved in 0.5 M sodium bicarbonatesolution (120 mL) and cooled to 0° C. Potassium iodide (15.0 g, 0.090mol) and iodine (7.66 g, 0.030 mol) were added followed by methylenechloride (40 mL). The solution was stirred at room temperatureovernight. After dilution with methylene chloride (100 mL), sodiumthiosulfate was added to quench the excess iodine. The organic layer wasseparated and washed with water (100 mL) and sodium chloride solution(100 mL). Drying over magnesium sulfate and evaporation of the solventgave S5-38 (5.44 g, 0.0137 mol, 91% yield).

¹H NMR: (CDCl₃) δ 1.86 (d of q, 1H, bridge —CH₂—), 2.47 (d of t, 1H,bridge —CH₂—), 2.83 (d of d, 1H, α-lactone carbonyl CH), 2.93 (bs, 1H,lactone bridgehead CH), 3.12 (d of d, 1H, α-benzyl ester CH), 3.29 (m,1H, bridgehead CH), 4.63 (d, 1H, α-lactone ester CH), 5.14 (d of d, 2H,CH₂Ph), 5.19 (d, 1H, iodo CH), 7.38 (m, 5H, C₆H₅).

Example 43 2-endo-Bezylcarboxy-bicylo[2.2.1]heptane-3,5-carbolactone(S5-39)

S5-38 (0.30 g, 0.75 mmol) was placed in DMSO under N₂, NaBH₄ (85 mg,2.25 mmol) added and the solution stirred at 85° C. for 2 h. The mixturewas cooled, diluted with water (50 mL) and extracted withdichloromethane (3×20 mL). The extracts were combined, washed with water(4×15 mL) and saturated aqueous NaCl (10 mL), dried, and evaporated togive 0.14 g (68%) of S5-39.

Example 445-endo-hydroxybicyclo[2.2.1]heptane-2-endo-benzyl-3-endo-methyldicarboxylate (S5-40)

Compound S5-39 is dissolved in methanol and sodium methoxide added withstirring. Removal of the solvent gives S5-40.

Example 45Bicyclo[2.2.1]heptane-2-endo-benzyl-3-endo-methyl-5-exo-(t-butoxycarbonyl)-aminodicarboxylate (S5-41)

In a one-pot reaction S5-40 is converted to the corresponding mesylatewith methanesulfonyl chloride, sodium azide added to displace themesylate to give exo-azide, which is followed by reduction with tributylphosphine to give the free amine, which is protected as the t-Bocderivative to give S5-41.

Example 46Bicyclo[2.2.1]heptane-2-endo-carboxy-3-exo-methyl-5-exo-(t-butoxycarbonyl)-aminocarboxylate (S5-42)

The benzyl ether protecting group is removed by catalytic hydrogenolysisof S5-41 with 10% Pd/C in methanol at room temperature for 6 hours.Filtration of the catalyst and removal of the solvent yields crudeS5-42.

Example 47Bicyclo[2.2.1]heptane-2-endo-carboxy-3-exo-methyl-5-exo-(t-butoxycarbonyl)-aminocarboxylate (S5-43)

Sodium is dissolved in methanol to generate sodium methoxide. S5-42 isadded and the mixture stirred at 62° C. for 16 hr. The mixture is cooledand acetic acid added with cooling to neutralize the excess sodiummethoxide. The mixture is diluted with water and extracted with ethylacetate. The extract is dried and evaporated to give S5-43.

Example 48Bicyclo[2.2.1]heptane-2-endo-benzyl-3-exo-methyl-5-exo-(t-butoxycarbonyl)aminodicarboxylate (S5-44)

Compound S5-43 is reacted with benzyl bromide and cesium carbonate intetrahydrofuran at room temperature to give benzyl ester S5-44, which isisolated by acid work-up of the crude reaction mixture.

Example 49Bicyclo[2.2.1]heptane-2-endo-benzyl-3-exo-carboxy-5-exo-(t-butoxycarbonyl)-aminocarboxylate (S5-45)

Compound S5-44 is dissolved in methanol and cooled to 0° C. under N₂. 2MNaOH (2 equivalents) is added dropwise, the mixture allowed to come toambient temperature and is stirred for 5 h. The solution is diluted withwater, acidified with 2M HCl and extracted with ethyl acetate. Theextract is washed with water, saturated aqueous NaCl, dried andevaporated to give S5-45.

Example 50Bicyclo[2.2.1]heptane-2-endo-benzyl-3-exo-(trimethylsilylethoxycarbonyl)amino-5-exo-(t-butoxycarbonyl)aminocarboxylate (S5-46)

To a solution of S5-45 in benzene is added triethylamine anddiphenylphosphoryl azide. The solution is refluxed for 24 hours and thencooled to room temperature. Trimethylsilylethanol is added and thesolution refluxed for 48 hours. The solution is partitioned betweenethyl acetate and 1M sodium bicarbonate. The organic layer is washedwith 1M sodium bicarbonate and dried over sodium sulfate. The solvent isevaporated under reduced pressure to give crude Curtius product S5-46.

Example 51endo-Bicyclo[2.2.1]hept-5-ene-2-(4-methoxy)benzylcarboxylate-3-carboxylicacid (S6-48)

Compound S3-19 and quinidine are suspended in equal amounts of tolueneand carbon tetrachloride and cooled to −55° C. p-Methoxybenzyl alcoholis added over 15 minutes and the solution stirred at −55° C. for 96hours. After removal of the solvents, the residue is partitioned betweenethyl acetate and 2 M hydrochloric acid. The organic layer is washedwith water and saturated aqueous sodium chloride. Drying over magnesiumsulfate and removal of the solvent gives S6-48.

Example 52endo-Bicyclo[2.2.1]hept-5-ene-2-(4-methoxy)benzyl-3-(trimethylsilylethoxy-carbonyl)aminocarboxylate (S6-49)

To a solution of S6-48 in benzene is added triethylamine anddiphenylphosphoryl azide. The solution is refluxed for 24 hours, cooledto room temperature, trimethylsilylethanol is added, and the solution isrefluxed for an additional 48 hours. The benzene solution is partitionedbetween ethyl acetate and 1 M sodium bicarbonate. The organic layers arecombined, washed with 1 M sodium bicarbonate, and dried with sodiumsulfate. The solvent is evaporated under reduced pressure to give crudeCurtius product S6-49.

Example 53Bicyclo[2.2.1]heptane-2-endo-(4-methoxy)benzyl-3-endo-(trimethylsilylethoxycarbonyl)amino-5-exo-methyl-6-exo-methyltricarboxylate (S6-50)

S6-49, copper(II) chloride, 10% Pd/C, and dry methanol are added to aflask with vigorous stirring. After degassing the suspension, the flaskis charged with carbon monoxide to a pressure just above 1 atm. Thepressure of carbon monoxide is maintained over 72 hours. The solids arefiltered off, and the crude reaction mixture worked up in the usual wayto afford S6-50.

Example 54Bicylo[2.2.1]heptane-2-endo-(4-methoxy)benzyl-3-endo-(trimethylsilylethoxycarbonyl)amino-5-exo-6-exo-dicarboxylicanhydride (S6-51)

S6-50, formic acid, and a catalytic amount of p-toluenesulfonic acid isheated at 90° C. overnight. Acetic anhydride is added to the reactionmixture, and it is refluxed for an additional 6 hours. Removal of thesolvents and washing with ether affords S6-51.

Example 55Bicyclo[2.2.1]heptane-2-endo-(4-methoxy)benzyl-3-endo-(trimethylsilylethoxycarbonyl)amino-5-exo-carboxy-6-exo-methyldicarboxylate (S6-52)

To a solution of S6-51 in equal amounts of toluene and carbontetrachloride is added quinine. The suspension is cooled to −65° C. andstirred for 1 hour. Three equivalents of methanol are added slowly over30 minutes. The suspension is stirred at −65° C. for 4 days followed byremoval of the solvents. The resulting white solid is partitionedbetween ethyl acetate and 2 M HCl, with S6-52 worked up from the organiclayer.

Example 56Bicyclo[2.2.1]heptane-2-endo-(4-methoxy)benzyl-3-endo-(trimethylsilylethoxycarbonyl)amino-5-exo-(trimethylsilylethoxycarbonyl)amino-6-exo-methyldicarboxylate (S6-53)

To a solution of S6-52 in benzene is added triethylamine anddiphenylphosphoryl azide. The solution is refluxed for 24 hours thencooled to room temperature. 2-Trimethylsilylethanol is added, and thesolution is refluxed for an additional 48 hours. The benzene solution ispartitioned between ethyl acetate and 1 M sodium bicarbonate. Theorganic layers are combined, washed with 1 M sodium bicarbonate, anddried with sodium sulfate. The solvent is evaporated under reducedpressure to give S6-53.

Example 57Bicyclo[2.2.1]heptane-2-exo-(4-methoxy)benzyl-3-endo-(trimethylsilylethoxycarbonyl)amino-5-exo-(trimethylsilylethoxycarbonyl)amino-6-endo-methyldicarboxylate (S6-54)

To a solution of S6-53 in tetrahydrofuran is carefully added potassiumtert-butoxide. The basic solution is refluxed for 24 hours followed byaddition of acetic acid. Standard extraction methods give the doubleepimerized product S6-54.

Example 58 Preparation of Hexamer

To 0.300 g (1R,2R)-(−)-trans-1,2-diaminocyclohexane (2.63 mmol) in 5 mLCH₂Cl₂ at 0° C. was added 0.600 g of 2,6-diformyl-4-bromophenol (2.62mmol) in 5 mL of CH₂Cl₂. The yellow solution was allowed to warm to roomtemperature and stirred for 48 hours. The reaction solution wasdecanted, and added to 150 mL of methanol. After standing for 30minutes, the yellow precipitate was collected, washed with methanol, andair-dried (0.580 g; 72% yield).

¹H NMR (400 MHz, CDCl₃) δ 14.31 (s, 3H, OH), 8.58 (s, 3H, CH═N), 8.19(s, 3H, CH═N), 7.88 (d, 3H, J=2.0 Hz, ArH), 7.27 (d, 3H, J=2.0 Hz, ArH),3.30-3.42 (m, 6H, CH₂—CH—N), 1.41-1.90 (m, 24H, aliphatic).

MS (FAB): Calcd for C₄₂H₄₆N₆O₃Br₃, 923.115. found 923.3 [M+H]⁺.

Example 59 Preparation of Hexamer

To 0.300 g (1R,2R)-(−)-trans-1,2-diaminocyclohexane (2.63 mmol) in 6 mLCH₂Cl₂ at 0° C. was added 0.826 g of2,6-diformyl-4-(1-dodec-1-yne)phenol (2.63 mmol) in 6 mL of CH₂Cl₂. Theorange solution was stirred at 0° C. for 1 hour and then allowed to warmto room temperature after which stirring was continued for 16 hours. Thereaction solution was decanted and added to 150 mL of methanol. A stickyyellow solid was obtained after decanting the methanol solution.Chromatographic cleanup of the residue gave a yellow powder.

¹H NMR (400 MHz, CDCl₃) δ 14.32 (s, 3H, OH), 8.62 (s, 3H, CH═N), 8.18(s, 3H, CH═N), 7.84 (d, 3H, J=2.0 Hz, ArH), 7.20 (d, 3H, J=2.0 Hz, ArH),3.30-3.42 (m, 6H, CH₂—CH—N), 2.25 (t, 6H, J=7.2 Hz, propargylic),1.20-1.83 (m, 72H, aliphatic), 0.85 (t, 9H, J=7.0 Hz, CH₃).

¹³C NMR (400 MHz, CDCl₃) δ 163.4, 161.8, 155.7, 136.9, 132.7, 123.9,119.0, 113.9, 88.7, 79.7, 75.5, 73.2, 33.6, 33.3, 32.2, 29.8, 29.7,29.6, 29.4, 29.2, 29.1, 24.6, 24.5, 22.9, 19.6, 14.4.

MS (FAB): Calcd for C₇₈H₁₀₉N₆O₃, 1177.856. found: 1177.8 [M+H]⁺.

Example 60 Preparation of Hexamer

To 0.240 g of 2,6-diformyl-4-(1-dodecene)phenol (0.76 mmol) in 10 mL ofbenzene was added a 10 mL benzene solution of(1R,2R)-(−)-trans-1,2-diaminocyclohexane (0.087 g, 0.76 mmol). Thesolution was stirred at room temperature for 48 hours shielded from thelight. The orange solution was taken to dryness and chromatographed(silica, 50/50 acetone/Et₂O) to give a yellow sticky solid (33% yield).

¹H NMR (400 MHz, CDCl₃) δ 14.12 (s, 3H, OH), 8.62 (s, 3H, CH═N), 8.40(s, 3H, CH═N), 7.82 (d, 3H, J=2.0 Hz, ArH), 7.28 (d, 3H, J=2.0 Hz, ArH),6.22 (d, 3H, vinyl), 6.05 (d, 3H, vinyl), 3.30-3.42 (m, 6H, CH₂—CH—N),1.04-1.98 (m, 87H, aliphatic).

MS (FAB): Calcd for C₇₈H₁₁₅N₆O₃, 1183.90. found: 1184.6 [M+H]⁺.

Example 61 Preparation of Tetramer

Preparation of hexamer:

Triethylamine (0.50 mL, 3.59 mmol) and(1R,2R)-(−)-trans-1,2-diaminocyclohexane (0.190 g, 1.66 mmol) werecombined in 150 mL EtOAc and purged with N₂ for 5 minutes. To thissolution was added 0.331 g isophthalolyl chloride (1.66 mmol) dissolvedin 100 mL EtOAc dropwise over six hours. The solution was filtered andthe filtrate taken to dryness. TLC (5% methanol/CH₂Cl₂) shows theproduct mixture to be primarily composed of two macrocycliccompositions. Chromatographic separation (silica, 5% methanol/CH₂Cl₂)gave the above tetramer (0.020 g, 5% yield) and hexamer (about 10%).

Tetramer:

¹H NMR (400 MHz, CDCl₃) δ 7.82 (s, 1H), 7.60 (br s, 2H), 7.45 (br s,2H), 7.18 (br s, 1H), 3.90 (br s, 2H), 2.22 (d, 2H), 1.85 (m, 4H), 1.41(m, 4H).

MS (ESI): Calcd for C₂₈H₃₃N₄O₄, 489.25. found 489.4 [M+H]⁺.

Hexamer:

MS (ESI): Calcd for C₄₂H₄₉N₆O₆, 733.37. found 733.5 [M+H]⁺.

Example 62 Preparation of Macrocyclic Modules from Benzene andCyclohexane Cyclic Synthons

To a 5 mL dichloromethane solution of 4-dodecyl-2,6-diformyl anisole (24mg; 0.072 mmol) was added a 5 mL dichloromethane solution of(1R,2R)-(−)-trans-1,2-diaminocyclohexane (8.5 mg; 0.074 mmol). Thissolution was stirred at room temperature for 16 hours and then added tothe top of a short silica column. Elution with diethyl ether and thenremoval of solvent led to the isolation of 22 mg of an off-white solid.Positive ion electrospray mass spectrometry demonstrated the presence ofthe tetramer (m/z 822, MH⁺), hexamer (m/z 1232, MH⁺), and the octamer(m/z 1643, MH⁺) in the off-white solid. Calculated molecular weightswere as follows: tetramer+H(C₅₄H₈₅N₄O₂, 821.67); hexamer+H(C₈₁H₁₂₇N₆O₃,1232.00); octamer+H(C₁₀₈H₁₆₉N₈O₄, 1643.33).

Example 63

Without intending to be bound by any one particular theory, one methodto approximate pore size of a macrocyclic module is quantum mechanical(QM) and molecular mechanical (MM) computations. In this example,macrocyclic modules having two types of synthons, “A” and “B,” were usedand all linkages between synthons were assumed to be the same. For thepurposes of QM and MM computations, the root mean square deviations inthe pore areas were computed over dynamic runs.

For QM, each module was first optimized using the MM+ force fieldapproach of Allinger (JACS, 1977, 99:8127) and Burkert, et al.,(Molecular Mechanics, ACS Monograph 177, 1982). They were thenre-optimized using the AM1 Hamiltonian (Dewar, et al., JACS, 1985,107:3903; Dewar, et al., JACS, 1986, 108:8075; Stewart, J. Comp. AidedMol. Design, 1990, 4:1). To verify the nature of the potential energysurface in the vicinity of the optimized structures, the associatedHessian matrices were computed using numerical double-differencing.

For MM, the OPLS-AA force field approach (Jorgensen, et al., JACS, 1996,118:11225) was used. For imine linkages, the dihedral angle was confinedto 180°±10°. The structures were minimized and equilibrated for onepicosecond using 0.5 femtosecond time steps. Then a 5 nanoseconddynamics run was carried out with a 1.5 femtosecond time step.Structures were saved every picosecond. The results are shown in Tables12 and 13.

Macrocyclic module pore areas derived from QM and MM computations forvarious linkages and macrocyclic module pore size are shown in Table 12.In Table 12, the macrocyclic modules had alternating synthons “A” and“B.” Synthon “A” is a benzene synthon coupled to linkages L at1,3-phenyl positions, and Synthon “B” is shown in the left-hand columnof the table.

TABLE 12 PORE AREAS FOR VARIOUS MACROCYCLIC MODULES (Å²) TETRAMERTETRAMER HEXAMER HEXAMER OCTAMER OCTAMER SYNTHON B QM MM QM MM QM MMtrans-1,2- imine (trans) Imine (trans) cyclohexane 14.3 Å² 13.2 ± 1.4 Å²trans-1,2- Acetylene cyclohexane 14.3 Å² trans-1,2- Amine Aminecyclohexane 23.1 Å² 13.9 ± 1.9 Å² trans-1,2- Amide Amide cyclohexane19.7 Å² 17.5 ± 2.0 Å² trans-1,2- Ester Ester cyclohexane 18.9 Å² 19.6 ±2.0 Å² Equatorial-1,3- imine (trans) Imine (trans) imine (trans) Imine(trans) cyclohexane 18.1 Å² 21.8 ± 1.6 Å² 66.2 Å² 74.5 ± 7.7 Å²Equatorial-1,3- Amine Amine cyclohexane 14.7 Å² 19.9 ± 2.6 Å²Equatorial-1,3- Amide Amide cyclohexane 24.8 Å² 21.7 ± 1.8 Å²Equatorial-1,3- Ester Ester cyclohexane 22.9 Å² 22.8 ± 2.4 Å²Equatorial-3- imine (trans) imine (trans) imine (trans) Imine (trans)imine (trans) Imine (trans) amino- oxygen-oxygen oxygen-oxygen 18.4 Å²21.0 ± 1.5 Å² 56.7 Å² 60.5⁺ − 8.3 Å² cyclohexene distance distance 2.481Å 3.7 ± .3 Å trans-1,2- imine (trans) Imine (trans) pyrrolidine 10.4 Å²9.2 ± 1.4 Å² Equatorial-1,3- imine (trans) Imine (trans) piperidene 19.2Å² 20.9 ± 1.1 Å² Endo-exo-1,2- imine (trans) Imine (trans)bicycloheptane 11.1 Å² 14.1 ± +−11 Å² Endo-endo-1,3- imine (trans) Imine(trans) bicycloheptane 18.8 Å² 20.7 ± 1.4 Å² Endo-exo-1,3- Imine Iminebicycloheptane 19.5 Å² 10.1 ± +4.9 Å² Equatorial-1,3- Amine Aminecyclohexane  9.8 Å² 9.9 ± 2.4 Å² Endo-endo-1,3- imine (trans) Imine(trans) bicyclooctene 18.9 Å² 21.6 ± 1.5 Å² Endo-exo-1,3- imine (trans)Imine (trans) bicyclooctene 15.6 Å² 18.7 ± 1.6 Å² Equatorial-3,9- imine(trans) Imine (trans) decalin 35.4 Å² 40.0 ± 2.2 Å²

Further macrocyclic module pore areas derived from QM and MMcomputations for various linkages and macrocyclic module pore size areshown in Table 13. In Table 13, the macrocyclic modules had alternatingsynthons “A” and “B.” In Table 13, Synthon “A” is a naphthalene synthoncoupled to linkages L at 2,7-naphthyl positions, and Synthon “B” isshown in the left-hand column of the table.

TABLE 13 PORE AREAS FOR VARIOUS MACROCYCLIC MODULES (Å²) HEXAMER HEXAMERSYNTHON B QM MM Trans-1,2- imine (trans) imine (trans) cyclohexane 23.5Å² 25.4 ± 4.9 Å² Endo-endo-1,3- imine (trans) imine (trans)bicycloheptane 30.1 Å² 30.0 ± 3.6 Å²

An example of the energy-minimized conformations of some hexamermacrocyclic modules having groups of substituents are shown in FIGS. 17Aand 17B. Referring to FIG. 17A, a Hexamer 1-h-(OH)₃ is shown having agroup of —OH substituents. Referring to FIG. 17B, a Hexamer 1-h-(OEt)₃is shown having a group of —OEt substituents. The differences in porestructure and area between these two examples, which also reflectconformational and flexibility differences, are evident. Thismacrocyclic module results in a composition which may be used toregulate pores. Selection of ethoxy synthon substituents over hydroxysynthon substituents for this hexamer composition is a method which maybe used for transporting selected species.

The pore size of macrocyclic modules was determined experimentally usinga voltage-clamped bilayer procedure. A quantity of a macrocyclic modulewas inserted into a lipid bilayer formed by phosphatidylcholine andphosphatidylethanolamine. On one side of the bilayer was placed asolution containing the cationic species to be tested. On the other sidewas a solution containing a reference cationic species known to be ableto pass through the pore of the macrocyclic module. Anions required forcharge balance were selected which could not pass through the pores ofthe macrocyclic module. When a positive electrical potential was appliedto the solution on the side of the lipid bilayer containing the testspecies, if the test species passed through the pores in the macrocyclicmodules, a current was detected. The voltage was then reversed to detectcurrent due to transport of the reference species through the pores,thereby confirming that the bilayer is a barrier to transport and thatthe pores of the macrocyclic modules provide transport of species.

Using the above technique, a hexameric macrocyclic module comprised of1R,2R-(−)-transdiaminocyclohexane and2,6-diformal-4-(1-dodec-1-ynyl)phenol synthons, having imine groups asthe linkages (the first module in Table 1) was tested for transport ofvarious ionic species. The results are shown in Table 14.

TABLE 14 VOLTAGE-CLAMPED BILAYER TEST FOR MACROCYCLIC MODULE PORE SIZECalculated van der Does ionic Calculated van der Waals radius of ionicspecies pass Waals radius of species with one through Ionic speciesionic species (Å) water shell (Å) pore? Na⁺ 1.0 2.2 Yes K⁺ 1.3 2.7 YesCa²⁺ 1.0 2.7 Yes NH₄ ⁺ 1.9 2.9 Yes Cs⁺ 1.7 3.0 Yes MeNH₃ ⁺ 2.0 3.0 YesEtNH₃ ⁺ 2.6 3.6 No NMe₄ ⁺ 2.6 3.6 No Aminoguani- 3.1 4.1 No dinium NEt₄⁺ 3.9 4.4 No Choline 3.8 4.8 No Glucosamine 4.2 5.2 No

The results in Table 14 show that the cut-off for passage through thepore in the selected module is a van der Waals radius of between 2.0 and2.6 Å. In Table 12, the QM and MM computed pore sizes are given asareas. Using the equation for area of a circle, A=πr², the computed areaof the pore in the first module of Table 12, 14.3 Å², gives a value forr of 2.13 Å. Ions having van der Waals radii of less than 2.13 Å wouldbe expected to traverse the pore and those with larger radii would not,and that is what was observed. CH₃NH₃ ⁺, having a radius of 2.0 Å,passed through the pore while CH₃CH₂NH₃ ⁺, with a radius of 2.6 Å, didnot. Without being held to a particular theory, and recognizing thatseveral factors influence pore transport, the observed ability ofhydrated ions to pass through the pore may be due to partial dehydrationof the species to enter the pore, transport of water molecules and ionsthrough the pore separately or with reduced interaction duringtransport, and re-coordination of water molecules and ions aftertransport. The details of pore structure, composition, and chemistry,the flexibility of the macrocyclic module, and other interactions mayaffect the transport process.

Example 64

Pore properties of 1,2-imine-linked and 1,2-amine-linked hexamermacrocyclic modules are illustrated in Table 15. Referring to Table 15,the bilayer clamp data indicates that the passage and exclusion ofcertain species through the pore of the modules correlates with thecomputational size of the pores. Further, these surprising data showthat a very small change in the placement of atoms and/or structuralfeatures can lead to a discrete change in transport properties and allowregulation of transport through the pore by variation of synthons andlinkages, among other factors.

TABLE 15 VOLTAGE-CLAMPED BILAYER TEST FOR MACROCYCLIC MODULE PORE SIZERadius of solute with H₂0 Hexamer 1a Hexamer 1jh Radius (radius of2^(nd) (1,2-imine) (1,2-amine) of hydration shell Radius = Radius =Solute species Solute in parentheses) 3.3 Å 3.9 Å Li⁺ 0.6 2.0 (5.6) NoYes Na⁺ 1.0 2.2 Yes Yes K⁺ 1.3 2.7 Yes Yes Ca²⁺ 1.0 2.7 Yes Yes Mg²⁺ 0.72.8 (5.5) No Yes NH₃ ⁺ 1.9 2.9 Yes Yes Cs⁺ 1.7 3 Yes Yes MeNH₃ ⁺ 2 3 YesYes EtNH₃ ⁺ 2.6 3.6 No Yes NMe₄ ⁺ 2.6 3.6 No Yes Aminoguani- 3.1 4.1 NoYes dine Choline 3.8 4.8 No Yes NEt₄ ⁺ 3.9 4.4 No No Glucosamine 4.2 5.2No No NPr₄ ⁺ — — — No

Example 65

The Langmuir isotherm and isobaric creep for hexamer 1a-Me are shown inFIGS. 18A and 18B, respectively.

The relative stability of the Langmuir film of Hexamer 1a-Me isillustrated by the isobaric creep data shown in FIG. 18B. The area ofthe film decreased by about 30% after about 30 min at 5 mN/m surfacepressure. The Langmuir isotherm and isobaric creep for Hexamer 1a-C15are shown in FIGS. 19A and 19B, respectively. The relative stability ofthe Langmuir film of Hexamer 1a-C15 is illustrated by the isobaric creepdata shown in FIG. 19B. The area of the film decreased by about 1-2%after about 30 min at 10 mN/m surface pressure, and by about 2% afterabout 60 min. The collapse pressure was about 18 mN/m for Hexamer1a-C15.

Example 66

Templated Imine Octamer. To a 3 neck 100 mL round bottomed flask withstirbar, fitted with condenser and addition funnel under argon,amphiphilic dialdehyde phenol 1 (500 mg, 1.16 mmol) was added. Next,Mg(NO₃)₂. 6H₂O (148 mg, 0.58 mmol) 2 and Mg(OAc)₂. 4H₂O (124 mg, 0.58mmol) were successively added. The flask was put under vacuo andbackfilled with argon 3 X. Anhydrous methanol was transferred to theflask via syringe under argon and the resulting suspension stirred. Themixture was then refluxed for 10 min affording a homogeneous solution.The reaction was allowed to cool to room temperature under positiveargon pressure. (1R,2R)-(−)-trans-1,2-diaminocyclohexane 4 was added tothe addition funnel followed by cannula transfer of anhydrous MeOH (11.6mL) under argon. The diamine/MeOH solution was added to the stirredhomogeneous metal template/dialdehyde solution drop wise over a periodof 1 h affording an orange oil. The addition funnel was replaced with aglass stopper and the mixture was refluxed for 3 days. The solvent wasremoved in vacuo affording a yellow crystalline solid that was usedwithout further purification.

Amine Octamer. To a 50 mL schlenk flask with stirbar under argon ImineOctamer (314 mg, 0.14 mmol) was added. Next anhydrous THF (15 mL) andMeOH (6.4 mL) were added via syringe under argon and the suspensionstirred at room temperature. To the homogeneous solution, NaBH₄ (136 mg,3.6 mmol) was added in portions and the mixture stirred at roomtemperature for 12 h. The solution was filtered, followed by addition of19.9 mL H₂O. The pH was adjusted to ca. 2 by addition of 4 M HCl, then6.8 mL of an ethylenediamine tetraaceticacid disodium salt dihydrate(0.13 M in H₂O) was added and the mixture stirred for 5 min. To thesolution, 2.0% ammonium hydroxide was added and stirring continued foran additional 5 min. The solution was extracted with ethyl acetate(3×100 mL) the organic layer separated, dried over Na₂SO₄ and thesolvent removed via rotoevaporation affording a pale yellow solid.Recrystallization from chloroform and hexanes afforded the AmineOctamer. Molecular weight was confirmed by ESIMS M+H=experimental=2058.7m/z, calcd=2058.7 m/z.

Example 67

Hexamer 1j. The two substrates, (−)—R,R-1,2-trans-diaminocyclohexane(0.462 mmol, 0.053 g) and 2,6-diformyl-4-hexadecyl benzylphenolcarboxylate (0.462 mmol, 0.200 g) were added to a 10 mL vial containinga magnetic stirbar followed by the addition of 2 mL of CH₂Cl₂. Theyellow solution was stirred at room temperature. After 24 h the reactionsolution was plugged through silica gel with diethyl ether, and thesolvent removed via roto-evaporation. (232 mg; 98% yield). ¹H NMR (400MHz, CDCl₃): δ 14.11 (s, 3H, OH), 8.67 (s, 3H, CH═N), 8.23 (s, 3H,CH═N), 7.70 (s, 3H, ArH), 7.11 (s, 3H, ArH), 4.05-3.90 (t, 6H, ³J=6.6Hz, CH₂C(O)OCH₂(CH₂)₁₄CH₃), 3.44 (s, 6H, CH₂C(O)OCH₂(CH₂)₁₄CH₃),3.30-3.42 (m, 6H, CH₂—CH—N), 1.21-1.90 (m, 108H, aliphatic) 0.92-0.86(t, 9H, ³J=6.6 Hz. ESIMS (+) Calcd for C₉₆H₁₅₁N₆O₉, 1533. Found: 1534[M+H]⁺.

Hexamer 1jh. To a 100 mL pear-shaped flask with magnetic stirbar underargon, Hexamer 1j (0.387 mmol, 0.594 g) was added and dissolved inTHF:MeOH (7:3, 28:12 mL, respectively). Next, NaBH₄ (2.32 mmol, 0.088 g)was added slowly in portions at room temperature for 6.5 h. The solventwas removed by roto-evaporation, the residue dissolved in 125 mL ethylacetate and washed 3×50 mL of H₂O. The organic layer was separated,dried over Na₂SO₄ and the solvent removed by roto-evaporation. Theresulting residue was recrystallized from CH₂Cl₂ and MeOH affording awhite solid (0.440 g; 74% yield). ¹H NMR (400 MHz, CDCl₃): δ 6.86 (s,6H, ArH), 4.10-4.00 (t, 6H, ³J=6.6 Hz, CH₂C(O)OCH₂(CH₂)₁₄CH₃), 3.87-3.69(dd, 6H, ³J=13.7 Hz, ³J (CNH)=42.4 Hz CH₂—CH—N), 3.43 (s, 6H,CH₂C(O)OCH₂(CH₂)₁₄CH₃), 2.40-2.28 (m, 6H, aliphatic), 2.15-1.95 (m, 6H,aliphatic), 1.75-1.60 (m, 6H, aliphatic), 1.60-1.55 (m, 6H, aliphatic)1.37-1.05 (m, 84H, aliphatic) 0.92-0.86 (t, 9H, ³J=6.8 Hz. ESIMS (+)Calcd for C₉₆H₁₆₃N₆O₉, 1544. Found: 1545 [M+H]⁺.

Example 68

Hexamer 1A-Me. A solution of2-hydroxy-5-methyl-1,3-benzenedicarboxaldehye (53 mg, 0.32 mmol) indichloromethane (0.6 mL) was added to a solution of(1R,2R)-(−)-1,2-diaminocyclohexane (37 mg, 0.32 mmol) in dichloromethane(0.5 mL). The mixture was stirred at ambient temperature for 16 h, addeddropwise to methanol (75 mL) and chilled (4° C.) for 4 h. Theprecipitate was collected to afford 71 mg (92%) of hexamer 1A-Me. ¹H NMR(CDCl₃): δ 13.88 (s, 3H, OH), 8.66 (s, 3H, ArCH═N), 8.19 (s, 3H,ArCH═N), 7.52 (d, 3H, J=2 Hz, Ar H), 6.86 (d, 3H, J=2 Hz, Ar H), 3.35(m, 6H, cyclohexane 1,2-H's), 2.03 (3, 9H, Me), 1.6-1.9 (m, 18H,cyclohexane 3,6-H₂ and 4_(eq),5_(eq)-H's), 1.45 (m, 6H, cyclohexane4_(ax),5_(ax)-H's); ¹³C NMR δ 63.67, 159.55, 156.38, 134.42, 129.75,127.13, 119.00, 75.68, 73.62, 33.68, 33.41, 24.65, 24.57, 20.22; ESI(+)MS m/e (%) 727 M+H (100); IR 1634 cm⁻¹.

Example 69

32.7 mg Hexamer 1jh (recrystallized times) was added to 30 mL dry THF.100 μL triethylamine and 100 μL acryloyl chloride (freshly distilled)were added subsequently to the THF mixture using Schlenk technique.Solution was stirred for 18 hrs in an acetone/dry ice bath. Afterremoval of solvent a white precipitate remained. The precipitate wasredissolved in CH₂Cl₂ and filtered through a fritted funnel. CH₂Cl₂solution was added to the separatory funnel and washed one time withwater followed by two brine (NaCl) washes. The CH₂Cl₂ solution was driedover MgSO₄ and then filtered to remove MgSO₄. A yellow precipitateremained after solvent removal. ¹H NMR (CDCl₃): δ −0.867-0.990 (3H),1.259 (21.8H), 1.39 (1.86H), 1.64 (12.7H), 2.8 (1.25H), 3.46-3.62(2.47H), 3.71 (0.89H), 3.99 (2.46H), 5.06 (0.71H), 5.31 (3.80H), 5.71(1.43H), 5.90 (0.78H), 6.2-6.4 (2.49H), 6.59 (0.80H), 6.78 (0.47H), 6.98(0.28H). FTIR-ATR: 3340, 2926 (—CH₂—), 2854 (—CH₂—), 1738 (EsterCarbonyl), 1649 and 1613 (Acrylate), 983 (═CH), 959 sh (═CH₂). ESI-MS:1978.5 (Hex1JhAC+8-AC), 1948.8 (Hex1JhAC+7-AC+Na⁺), 1923.3(Hex1JhAC+7-AC), 1867.6 (Hex1JhAC+6-AC), 1842.6, 1759.7 (Hex1JhAC+4-AC).

1. A method for filtration comprising contacting a nanofilm with a fluidto provide a filtered fluid, wherein said nanofilm comprises amphiphilicmacrocyclic modules, wherein at least one of said amphiphilicmacrocyclic modules comprises three to about twenty-four cyclic synthonscoupled into a closed ring by one or more coupling linkages between saidcyclic synthons including at least one linkage other than —CH₂—; whereinsaid fluid comprises one or more components for which said nanofilm haslow permeability and one or more components for which said nanofilm ispermeable; and wherein said nanofilm retains at least some of said oneor more low permeability components and wherein at least some of saidpermeable components traverse the nanofilm.
 2. The method of claim 1,wherein said fluid is selected from at least one gas, at least oneliquid, and mixtures thereof.
 3. The method of claim 1, wherein at leastsome of said amphiphilic macrocyclic modules define a pore having aradius of about 0.5 Å to about 5 Å.
 4. The method of claim 3, whereinthe pore has a radius of about 2.13 Å.
 5. The method of claim 3, whereinthe pore has a radius of about 3.3 Å.
 6. The method of claim 3, whereinthe pore has a radius of about 3.9 Å.
 7. The method of claim 3, whereinsaid one or more low permeability components and said one or morepermeable components may be solvated.
 8. The method of claim 3, whereinsaid one or more low permeability components and said one or morepermeable components may be non-solvated.
 9. The method of claim 7,wherein said one or more solvated low permeability components have aradius greater than that of the pore.
 10. The method of claim 8, whereinsaid one or more non-solvated low permeability components have a radiusgreater than that of the pore.
 11. The method of claim 9, wherein saidone or more solvated low permeability components will not traverse thepore.
 12. The method of claim 10, wherein said one or more non-solvatedlow permeability components will not traverse the pore.
 13. The methodof claim 1, wherein said one or more low permeability components have amolecular weight greater than about 50 Da.
 14. The method of claim 1,wherein said one or more low permeability components have a molecularweight greater than about 100 Da.
 15. The method of claim 1, whereinsaid one or more low permeability components have a molecular weightgreater than about 400 Da.
 16. The method of claim 1, wherein said oneor more low permeability components have a molecular weight greater thanabout 600 Da.
 17. The method of claim 1, wherein said one or more lowpermeability components have a molecular weight greater than about 800Da.
 18. The method of claim 1, wherein said one or more low permeabilitycomponents have a molecular weight greater than about 1 kDa.
 19. Themethod of claim 1 wherein said one or more low permeability componentshave a clearance through the nanofilm of less than about 10%.
 20. Themethod of claim 1 wherein said one or more low permeability componentshave a clearance through the nanofilm of less than about 20%.
 21. Themethod of claim 1 wherein said one or more low permeability componentshave a clearance through the nanofilm of less than about 30%.
 22. Themethod of claim 1 wherein said nanofilm is permeable to water.
 23. Themethod of claim 22 wherein said one or more low permeability componentscomprise one or more of immunoglobulin G, albumin, β₂-Microglobulin,myoglobin, ovalbumin, glucose, urea, creatinine, Li⁺, Ca²⁺, Mg²⁺, andviruses.
 24. The method of claim 22 wherein said nanofilm is permeableto Cl⁻.
 25. The method of claim 22 wherein said nanofilm is permeable toK⁺.
 26. The method of claim 22 wherein said one or more low permeabilitycomponents comprise Na⁺.
 27. The method of claim 25 wherein said one ormore low permeability components comprise Na⁺.
 28. The method of claim22 wherein said nanofilm is permeable to Na⁺.
 29. The method of claim 22wherein said one or more low permeability components comprise K⁺. 30.The method of claim 28 wherein said one or more low permeabilitycomponents comprise K⁺.
 31. The method of claim 28 wherein said nanofilmis permeable to K⁺and glucose.
 32. The method of claim 22, wherein saidnanofilm is permeable to Na⁺, K⁺, hydrogen phosphate and dihydrogenphosphate.
 33. The method of claim 1, wherein at least one of saidamphiphilic macrocyclic modules is coupled to at least one secondamphiphilic macrocyclic module through at least one reactive functionalgroup.
 34. The method of claim 1, wherein said amphiphilic macrocyclicmodules are independently selected from Hexamer 1a, Hexamer 1dh, Hexamer3j- amine, Hexamer 1jh, Hexamer 1jh-AC, Hexamer 2j-amine/ester, Hexamer1dh-acryl, Octamer 5jh-aspartic, and Octamer 4jh-acryl.
 35. The methodof claim 1 wherein said nanofilm further comprises one or more surfaceattachment groups.
 36. The method of claim 35 wherein said one or moresurface attachment groups are independently selected from amino,hydroxyl, halo, thiol, alkynyl, magnesium halo, aldehyde, —CH═C(CH₃)₂,vinyl, —(CH═CH)—CH═CH₂, —OC(O)CH(CH₃)₂, —OC(O)CH═CH₂, —N(C(O)CH═CH₂,carboxylate, isocyanate, epoxide, and streptavidin.
 37. The method ofclaim 1 wherein at least one of said amphiphilic macrocyclic modules hasone or more hydrophobic tails that are cleavable from said at least oneof said macrocyclic modules by at least one method chosen from chemical,thermal, photochemical, electrochemical, and irradiative.
 38. The methodof claim 1 wherein at least one of said amphiphilic macrocyclic moduleshas one or more hydrophilic groups.
 39. The method of claim 38 whereinsaid one or more hydrophilic groups are independently selected fromhydroxyl, methoxy, phenol, carboxylic acids and salts thereof, methyl-ethyl-, and vinyl-esters of carboxylic acids, amides, amino, cyano,ammonium salts, sulfonium salts, phosphonium salts, polyethyleneglycols, epoxy groups, acrylates, sulfonamides, nitro, —OP(O)(OCH₂CH₂N⁺RR′R″)O⁻, guanidinium, aminate, acrylamide, pyridinium, andpiperidine, wherein R, R′, and R″ are each independently selected from Hand alkyl.
 40. The method of claim 1 wherein at least one of saidamphiphilic macrocyclic modules comprises one or more functional groupsto impart amphiphilic character wherein said one or more functionalgroups are attached to said amphiphilic macrocyclic modules via alinkage chosen from carboxylate and amide.
 41. The method of claim 40wherein said one or more functional groups to impart amphiphiliccharacter are independently selected from

and

wherein m is seven to twenty-seven.
 42. The method of claim 33 whereinsaid at least one of said amphiphilic macrocyclic modules and said atleast one second amphiphilic macrocyclic module are coupled to at leastone linker molecule.
 43. The method of claim 42 wherein said at leastone linker molecule is selected from

and mixtures thereof; wherein m is one to ten, n is one to six, each Ris independently chosen from H and CH₃, each R′ is independently chosenfrom —(CH₂)_(n)—and phenylene, each R″ is independently chosen from—(CH₂)_(n)—, polyethylene glycol (PEG), and polypropylene glycol (PPG),and each X is independently chosen from Br, Cl, and I.
 44. The method ofclaim 1 wherein one or more of said amphiphilic macrocyclic modules haveat least one reactive functional group independently selected from Table2.